Bicyclic heteroarylaminoalkyl phenyl derivatives as pi3k inhibitors

ABSTRACT

This application relates to derivatives of Formula I: 
     
       
         
         
             
             
         
       
     
     and pharmaceutically acceptable salts thereof, which are inhibitors of PI3K, and compositions and methods of treatment related thereto.

This application is a continuation of U.S. Ser. No. 16/112,174, filedAug. 24, 2018, which is a divisional of U.S. Ser. No. 14/735,438, filedJun. 10, 2015, which claims the benefit of priority of Ser. No.62/010,760, filed Jun. 11, 2014, each of which is incorporated herein byreference in its entirety.

TECHNICAL FIELD

This application relates to bicyclic heteroarylaminoalkyl phenylderivatives, which are inhibitors of PI3K, and compositions and methodsof treatment related thereto.

BACKGROUND

The phosphoinositide 3-kinases (PI3Ks) belong to a large family of lipidsignaling kinases that phosphorylate phosphoinositides at the D3position of the inositol ring (Cantley, Science, 2002, 296(5573):1655-7). PI3Ks are divided into three classes (class I, II, andIII) according to their structure, regulation and substrate specificity.Class I PI3Ks, which include PI3Kα, PI3Kβ, PI3Kγ, and PI3Kδ, are afamily of dual specificity lipid and protein kinases that catalyze thephosphorylation of phosphatidylinosito-4,5-bisphosphate (PIP₂) givingrise to phosphatidylinosito-3,4,5-trisphosphate (PIP₃). PIP₃ functionsas a second messenger that controls a number of cellular processes,including growth, survival, adhesion and migration. All four class IPI3K isoforms exist as heterodimers composed of a catalytic subunit(p110) and a tightly associated regulatory subunit that controls theirexpression, activation, and subcellular localization. PI3Kα, PI3Kβ, andPI3Kδ associate with a regulatory subunit known as p85 and are activatedby growth factors and cytokines through a tyrosine kinase-dependentmechanism (Jimenez, et al., J Biol Chem., 2002, 277(44):41556-62)whereas PI3Kγ associates with two regulatory subunits (p101 and p84) andits activation is driven by the activation of G-protein-coupledreceptors (Brock, et al., J Cell Biol., 2003, 160(1):89-99). PI3Kα andPI3Kβ are ubiquitously expressed. In contrast, PI3Kγ and PI3Kδ arepredominantly expressed in leukocytes (Vanhaesebroeck, et al., TrendsBiochem Sci., 2005, 30(4):194-204).

The differential tissue distribution of the PI3K isoforms factors intheir distinct biological functions. Genetic ablation of either PI3Kα orPI3Kβ results in embryonic lethality, indicating that PI3Kα and PI3Kβhave essential and non-redundant functions, at least during development(Vanhaesebroeck, et al., 2005). In contrast, mice which lack PI3Kγ andPI3Kδ are viable, fertile and have a normal life span although they showan altered immune system. PI3Kγ deficiency leads to impaired recruitmentof macrophages and neutrophils to sites of inflammation as well asimpaired T cell activation (Sasaki, et al., Science, 2000,287(5455):1040-6). PI3δ-mutant mice have specific defects in B cellsignaling that lead to impaired B cell development and reduced antibodyresponses after antigen stimulation (Clayton, et al., J Exp Med. 2002,196(6):753-63; Jou, et al., Mol Cell Biol. 2002, 22(24):8580-91;Okkenhaug, et al., Science, 2002, 297(5583):1031-4).

The phenotypes of the PI3Kγ and PI3Kδ-mutant mice suggest that theseenzymes may play a role in inflammation and other immune-based diseasesand this is borne out in preclinical models. PI3Kγ-mutant mice arelargely protected from disease in mouse models of rheumatoid arthritis(RA) and asthma (Camps, et al., Nat Med. 2005, 11(9):936-43; Thomas, etal., Eur. J. Immunol. 2005, 35(4):1283-91). In addition, treatment ofwild-type mice with a selective inhibitor of PI3Kγ was shown to reduceglomerulonephritis and prolong survival in the MRL-lpr model of systemiclupus nephritis (SLE) and to suppress joint inflammation and damage inmodels of RA (Barber, et al., Nat Med. 2005, 11(9):933-5; Camps, et al.,2005). Similarly, both PI3Kδ-mutant mice and wild-type mice treated witha selective inhibitor of PI3Kδ have been shown to have attenuatedallergic airway inflammation and hyper-responsiveness in a mouse modelof asthma (Ali, et al., Nature. 2004, 431(7011):1007-11; Lee, et al.,FASEB J. 2006, 20(3):455-65) and to have attenuated disease in a modelof RA (Randis, et al., Eur. J. Immunol., 2008, 38(5):1215-24).

B cell proliferation has shown to play a major role in the developmentof inflammatory autoimmune diseases (Puri, Frontiers in Immunology(2012), 3(256), 1-16; Walsh, Kidney International (2007) 72, 676-682).For example, B cells support T-cell autoreactivity, an importantcomponent of inflammatory autoimmune dieases. Once activated andmatured, B cells can traffic to sites of inflammation and recruitinflammatory cells or differentiate to plasmablasts. Thus, activity ofB-cells can be affected by targeting B-cell stimulatory cytokines,B-cell surface receptors, or via B-cell depletion. Rituximab—an IgG1 κmouse/human chimeric monoclonal antibody directed against the B-cellsurface receptor CD20—has been shown to deplete CD20+ B cells. Use ofrituximab has been shown to have efficacy in treating idiopathicthrombocytopenic purpura, autoimmune hemolytic anemia, or vasculitis.For example, treatment with rituximab resulted in remission of thedisease in patients suffering from anti-neutrophil cytoplasm antibodyassociated (ANCA) systemic vasculitis (AASV) with demonstratedperipheral B-cell depletion (Walsh, 2007; Lovric, Nephrol DialTransplant (2009) 24: 179-185). Similarly, a complete response wasreported in one-third to two-thirds of patients having mixedcryoglobulinemia vasculitis after treatment with rituximab, includingpatients who presented with a severe form of vasculitis that wasresistant or intolerant to other treatments (Cacoub, Ann Rheum Dis 2008;67:283-287). Similarly, rituximab has been shown to have efficacy intreating patients with idiopathic thrombocytopenic purpura (or immunethrombocytopenic purpura) (Garvey, British Journal of Haematology,(2008) 141, 149-169; Godeau, Blood (2008), 112(4), 999-1004; Medeo,European Journal of Haematology, (2008) 81, 165-169) and autoimmunehemolytic anemia (Garvey, British Journal of Haematology, (2008) 141,149-169).

PI3Kδ signaling has been tied to B cell survival, migration, andactivation (Puri, Frontiers in Immunology, 2012, 3(256), 1-16, at pages1-5; and Clayton, J Exp Med, 2002, 196(6):753-63). For example, PI3Kδ isrequired for antigen-dependent B-cell activation driven by B cellreceptor. By blocking B-cell adhesion, survival, activation, andproliferation, PI3Kδ inhibition can impair the ability of B cells toactivate T cells, preventing their activation and reducing secreation ofautoantibodies and pro-inflammatory cytokines. Hence, by their abilityto inhibit B cell activation, PI3Kδ inhibitors would be expected totreat B cell mediated diseases that were treatable by similar methodssuch as B cell depletion by rituximab. Indeed, PI3Kδ inhibitors havebeen shown to be useful mouse models of various autoimmune diseases thatare also treatable by rituximab such as arthritis (Puri (2012)).Further, innate-like B cells, which are linked to autoimmunity aresensitive to PI3Kδ activity, as MZ and B-1 cells are nearly absent inmice lacking the p110δ gene (Puri (2012). PI3Kδ inhibitors can reducetrafficking of and activation of MZ and B-1 cells, which are implicatedin autoimmune diseases.

In addition to their potential role in inflammatory diseases, all fourclass I PI3K isoforms may play a role in cancer. The gene encoding p110αis mutated frequently in common cancers, including breast, prostate,colon and endometrial (Samuels, et al., Science, 2004, 304(5670):554;Samuels, et al., Curr Opin Oncol. 2006, 18(1):77-82). Eighty percent ofthese mutations are represented by one of three amino acid substitutionsin the helical or kinase domains of the enzyme and lead to a significantupregulation of kinase activity resulting in oncogenic transformation incell culture and in animal models (Kang, et al., Proc Natl Acad Sci USA.2005, 102(3):802-7; Bader, et al., Proc Natl Acad Sci USA. 2006,103(5):1475-9). No such mutations have been identified in the other PI3Kisoforms although there is evidence that they can contribute to thedevelopment and progression of malignancies. Consistent overexpressionof PI3Kδ is observed in acute myeloblastic leukemia (Sujobert, et al.,Blood, 2005, 106(3):1063-6) and inhibitors of PI3Kδ can prevent thegrowth of leukemic cells (Billottet, et al., Oncogene. 2006,25(50):6648-59). Elevated expression of PI3Kγ is seen in chronic myeloidleukemia (Hickey, et al., J Biol Chem. 2006, 281(5):2441-50).Alterations in expression of PI3Kβ, PI3Kγ and PI3Kδ have also beenobserved in cancers of the brain, colon and bladder (Benistant, et al.,Oncogene, 2000, 19(44):5083-90; Mizoguchi, et al., Brain Pathol. 2004,14(4):372-7; Knobbe, et al., Neuropathol Appl Neurobiol. 2005,31(5):486-90). Further, these isoforms have all been shown to beoncogenic in cell culture (Kang, et al., 2006).

For these reasons, there is a need to develop new PI3K inhibitors thatcan be used inflammatory disorders, autoimmune diseases and cancer. Thisinvention is directed to this need and others.

SUMMARY

The present application provides, inter alia, a compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein the variables aredefined infra.

The present application further provides compositions comprising acompound of the invention, or a pharmaceutically acceptable saltthereof, and at least one pharmaceutically acceptable carrier.

The present application also provides methods of modulating an activityof a PI3K kinase, comprising contacting the kinase with a compound ofthe invention, or a pharmaceutically acceptable salt thereof.

The present application further provides methods of treating a diseasein a patient, wherein said disease is associated with abnormalexpression or activity of a PI3K kinase, comprising administering tosaid patient a therapeutically effective amount of a compound of theinvention, or a pharmaceutically acceptable salt thereof.

The present application further provides methods of treating animmune-based disease in a patient, comprising administering to saidpatient a therapeutically effective amount of a compound of theinvention, or a pharmaceutically acceptable salt thereof.

The present application also provides methods of treating a cancer in apatient, comprising administering to said patient a therapeuticallyeffective amount of a compound of the invention, or a pharmaceuticallyacceptable salt thereof.

The present application further provides methods of treating a lungdisease in a patient, comprising administering to said patient atherapeutically effective amount of a compound of the invention, or apharmaceutically acceptable salt thereof.

The present application also provides a compound of the invention, or apharmaceutically acceptable salt thereof, for use in any of the methodsdescribed herein.

The present application further provides use of a compound, or apharmaceutically acceptable salt thereof, for the manufacture of amedicament for use in any of the methods described herein.

DETAILED DESCRIPTION

The present application provides, inter alia, a compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein:

Ar is

X is N or CR¹⁰;

X¹ is O or S;

R¹ is alkyl;

R² is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkyl, C₁₋₆ haloalkoxy,phenyl, or 5-6 membered heteroaryl; wherein said phenyl and 5-6 memberedheteroaryl are each optionally substituted by 1, 2, 3, or 4 substituentsindependently selected from halo, OH, CN, C₁₋₄ alkyl, C₁₋₄ alkoxy, andC₁₋₄ haloalkoxy;

R³ is Cy, —(C₁₋₃ alkylene)—Cy, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₁₋₆ haloalkyl, C(═O)R^(b), C(═O)NR^(c)R^(d), C(═O)OR^(a), NR^(c)R^(d),NR^(c)C(═O)R^(a), NR^(c)C(═O)OR^(b), NR^(c)C(═O)NR^(c)R^(d),NR^(c)S(═O)₂R^(b), or NR^(c)S(═O)₂NR^(c)R^(d); wherein said C₁₋₆ alkyl,C₂₋₆ alkenyl, and C₂₋₆ alkynyl are each optionally substituted by 1, 2,or 3 independently selected R^(3a) groups;

provided that either (i) R² is phenyl or 5-6 membered heteroaryl,wherein said phenyl and 5-6 membered heteroaryl are each optionallysubstituted by 1, 2, 3, or 4 substituents independently selected fromhalo, OH, CN, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkoxy; or (ii) R³is Cy or —(C₁₋₃ alkylene)-Cy;

R⁴ is H, halo, OH, CN, C₁₋₄ alkyl, C₁₋₄ haloalkyl, C₁₋₄ alkoxy, or C₁₋₄haloalkoxy;

R⁵ is halo, OH, CN, C₁₋₄ alkyl, C₁₋₄ haloalkyl, C₁₋₄ alkoxy, C₁₋₄haloalkoxy, or cyclopropyl;

R⁶ is H, halo, CN, or C₁₋₄ alkyl;

each R⁸ is independently selected from OH, NO₂,CN, halo, C₁₋₃ alkyl,C₂₋₃ alkenyl, C₂₋₃ alkynyl, C₁₋₃ haloalkyl, cyano-C₁₋₃ alkyl, HO—C₁₋₃alkyl, C₁₋₃ alkoxy-C₁₋₃ alkyl, C₃₋₇ cycloalkyl, C₁₋₃ alkoxy, C₁₋₃haloalkoxy, amino, C₁₋₃ alkylamino, di(C₁₋₃ alkyl)amino, thio, C₁₋₃alkylthio, C₁₋₃ alkylsulfinyl, C₁₋₃ alkylsulfonyl, carbamyl, C₁₋₃alkylcarbamyl, di(C₁₋₃ alkyl)carbamyl, carboxy, C₁₋₃ alkylcarbonyl, C₁₋₄alkoxycarbonyl, C₁₋₃ alkylcarbonylamino, C₁₋₃ alkylsulfonylamino,aminosulfonyl, C₁₋₃ alkylaminosulfonyl, alkyl)aminosulfonyl,aminosulfonylamino, C₁₋₃ alkylaminosulfonylamino,alkyl)aminosulfonylamino, aminocarbonylamino, C₁₋₃alkylaminocarbonylamino, and di(C₁₋₃ alkyl)aminocarbonylamino;

each R¹⁰, R¹³, and R¹⁴ is independently hydrogen, OH, NO₂,CN, halo, oxo,C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄ haloalkyl, cyano-C₁₋₄alkyl, HO—C₁₋₃ alkyl, C₁₋₃ alkoxy-C₁₋₃ alkyl, C₃₋₇ cycloalkyl, alkoxy,C₁₋₃ haloalkoxy, amino, alkylamino, di(C₁₋₃ alkyl)amino, thio,alkylthio, alkylsulfinyl, alkylsulfonyl, carbamyl, C₁₋₃ alkylcarbamyl,di(C₁₋₃ alkyl)carbamyl, carboxy, C₁₋₃ alkylcarbonyl, C₁₋₄alkoxycarbonyl, C₁₋₃ alkylcarbonylamino, C₁₋₃ alkylsulfonylamino,aminosulfonyl, C₁₋₃ alkylaminosulfonyl, di(C₁₋₃ alkyl)aminosulfonyl,aminosulfonylamino, C₁₋₃ alkylaminosulfonylamino, di(C₁₋₃alkyl)aminosulfonylamino, aminocarbonylamino, C₁₋₃alkylaminocarbonylamino, di(C₁₋₃ alkyl)aminocarbonylamino, C₃₋₇cycloalkyl, 4-7 membered heterocycloalkyl, phenyl, phenyl-C₁₋₃-alkyl,5-6 membered heteroaryl, 5-6 membered heteroaryl-C₁₋₃-alkyl, and 4-7membered heterocycloalkyloxy; and

each R¹¹, R¹², and R¹⁵ is independently hydrogen, C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alkynyl, C₁₋₄ haloalkyl, cyano-C₁₋₄ alkyl, HO—C₁₋₃ alkyl,C₁₋₃ alkoxy-C₁₋₃ alkyl, C₃₋₇ cycloalkyl, C₁₋₃ alkoxy, C₁₋₃ haloalkoxy,amino, C₁₋₃ alkylamino, di(C₁₋₃alkyl)amino, thio, C₁₋₃ alkylthio, C₁₋₃alkylsulfinyl, C₁₋₃ alkylsulfonyl, carbamyl, C₁₋₃ alkylcarbamyl, di(C₁₋₃alkyl)carbamyl, carboxy, C₁₋₃ alkylcarbonyl, C₁₋₄ alkoxycarbonyl, C₁₋₃alkylcarbonylamino, C₁₋₃ alkylsulfonylamino, aminosulfonyl, C₁₋₃alkylaminosulfonyl, di(C₁₋₃ alkyl)aminosulfonyl, aminosulfonylamino,C₁₋₃ alkylaminosulfonylamino, di(C₁₋₃ alkyl)aminosulfonylamino,aminocarbonylamino, C₁₋₃ alkylaminocarbonylamino, di(C₁₋₃alkyl)aminocarbonylamino, C₃₋₇ cycloalkyl, 4-7 memberedheterocycloalkyl, phenyl, aryl-C₁₋₃-alkyl, 5-6 membered heteroaryl, 5-6membered heteroaryl-C₁₋₃-alkyl, and 4-7 membered heterocycloalkyloxy;

each R^(a)R^(c), and R^(d) is independently selected from H, C₁₋₆ alkyl,C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, and Cy; wherein said C₁₋₆alkyl, C₂₋₆ alkenyl, and C₂₋₆ alkynyl are each optionally substitutedwith 1, 2, or 3 independently selected R^(3b) groups;

each R^(b) is independently selected from C₁₋₆ alkyl, C₁₋₆ haloalkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, and Cy; wherein said C₁₋₆ alkyl, C₂₋₆alkenyl, and C₂₋₆ alkynyl are each optionally substituted with 1, 2, or3 independently selected R^(3b) groups;

or R^(c) and R^(d) together with the N atom to which they are attachedform a 4-, 5-, 6-, or 7 membered heterocycloalkyl group, which isoptionally substituted with —OH or C₁₋₃ alkyl;

each Cy is independently selected from C₃₋₇ cycloalkyl, 4-6 memberedheterocycloalkyl, phenyl, naphthyl, and 5-6 membered heteroaryl, each ofwhich is optionally substituted with 1, 2, or 3 independently selectedR^(3b) groups;

each R^(3a) is independently selected from halo, CN, NO₂,C₁₋₆ alkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, OR^(a1), SR^(a1),C(═O)R^(b1), C(═O)NR^(c1)R^(d1), C(═O)OR^(a1), OC(═O)R^(b1),OC(═O)NR^(c1)R^(d1), NR^(c1)R^(d1), NR^(c1)C(═O)R^(a1),NR^(c1)C(═O)OR^(b1), NR^(c1)C(═O)NR^(c1)R^(d1), C(═NR^(c))R^(b1),C(═NR^(e))NR^(c1)R^(d1), NR^(c1)C(═NR^(e))NR^(c1)R^(d1),NR^(c1)S(═O)R^(b1), NR^(c1)S(═O)₂NR^(c1)R^(d1), S(═O)₂R^(b1), andS(═O)₂NR^(c1)R^(d1); wherein said C₁₋₆ alkyl, C₂₋₆ alkenyl, and C₂₋₆alkynyl are each optionally substituted with 1, 2, or 3 independentlyselected R⁸ groups;

each R^(3b), is independently selected from Cy¹, —(C₁₋₃ alkylene)-Cy¹,halo, CN, NO₂, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl,OR^(a1), SR^(a1), C(═O)R^(b1), C(═O)NR^(c1)R^(d1); C(═O)OR^(a1),OC(═O)R^(b1), OC(═O)NR^(c1)R^(d1), NR^(c1)R^(d1), NR^(c1)C(═O)R^(a1),NR^(c1)C(═O)OR^(b1), NR^(c1)C(═O)NR^(c1)R^(d1), C(═NR^(e))R^(b1),C(═NR^(e))NR^(c1)R^(d1), NR^(c1)C(═NR^(e))NR^(c1)R^(d1),NR^(c1)S(═O)R^(b1), NR^(c1)S(═O)₂NR^(c1)R^(d1), S(═O)R^(b1),S(═O)₂R^(b1), and S(═O)₂NR^(c1)R^(d1); wherein said C₁₋₆ alkyl, C₂₋₆alkenyl, and C₂₋₆ alkynyl are each optionally substituted with 1, 2, or3 independently selected R⁸ groups;

each Cy¹ is independently selected from C₃₋₇ cycloalkyl, 4-7 memberedheterocycloalkyl, phenyl, and 5-6 membered heteroaryl, each of which isoptionally substituted with 1, 2, 3, or 4 independently selected R⁸groups;

each R^(a1), R^(c1), and R^(d1) is independently selected from H, C₁₋₆alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₇ cycloalkyl, 4-7membered heterocycloalkyl, phenyl, and 5-6 membered heteroaryl; whereinsaid C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₇ cycloalkyl, 4-7membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl are eachoptionally substituted with 1, 2, or 3 independently selected R⁸ groups;and

each R^(b1) is independently selected from C₁₋₆ alkyl, C₁₋₆ haloalkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₇ cycloalkyl, 4-7 memberedheterocycloalkyl, phenyl, and 5-6 membered heteroaryl; wherein said C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₇ cycloalkyl, 4-7 memberedheterocycloalkyl, phenyl and 5-6 membered heteroaryl are each optionallysubstituted with 1, 2, or 3 independently selected R⁸ groups;

or R^(c1) and R^(d1) together with the N atom to which they are attachedform a 4-, 5-, 6-, or 7 membered heterocycloalkyl group, which isoptionally substituted with —OH or C₁₋₃ alkyl.

In some embodiments, R⁶ is H.

In some embodiments, R¹ is methyl.

In some embodiments, R² is C₁₋₆ alkoxy or phenyl; wherein said phenyl isoptionally substituted by 1, 2, 3, or 4 substituents independentlyselected from halo, OH, CN, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄haloalkoxy.

In some embodiments, R² is C₁₋₄ alkoxy.

In some embodiments, R² is methoxy or ethoxy.

In some embodiments, R² is phenyl; wherein said phenyl is optionallysubstituted by 1, 2, 3, or 4 independently selected halo groups.

In some embodiments, wherein R² is 3,5-difluorophenyl.

In some embodiments, R³ is Cy, C(═O)R^(b), C(═O)NR^(c)R^(d), orC(═O)OR^(a).

In some embodiments, R³ is Cy or C(═O)NR^(c)R^(d).

In some embodiments, R³ is Cy.

In some embodiments, R³ is C(═O)NR^(c)R^(d).

In some embodiments, R³ is a 4-6 membered heterocycloalkyl or 5-6membered heteroaryl, each of which is optionally substituted by 1 or 2independently selected R^(3b) groups.

In some embodiments, R³ is:

In some embodiments, R³ is pyridine optionally substituted by 1 or 2independently selected R^(3b) groups.

In some embodiments, R³ is:

In some embodiments, R³ is C(═O)NH(CH₂CH₃).

In some embodiments, each R^(3b) is independently selected from halo,CN, NO₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl, OR^(a1), C(═O)R^(b1),C(═O)NR^(c1)R^(d1), C(═O)OR^(a1), NR^(c1)R^(d1), S(═O)₂R^(b1), andS(═O)₂NR^(c1)R^(d1); wherein said C₁₋₆ alkyl is optionally substitutedwith 1, 2, or 3 independently selected R⁸ groups.

In some embodiments, each R^(3b) is independently selected from halo,CN, C₁₋₆ alkyl, C₁₋₆ haloalkyl, hydroxy, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,amino, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, C₁₋₆ alkylsulfonyl,carbamyl, C₁₋₆ alkylcarbamyl, and di(C₁₋₆ alkyl)carbamyl.

In some embodiments, each R^(3b) is C₁₋₆ alkylamino.

In some embodiments, each R^(3b) is di(C₁₋₆ alkyl)carbamyl.

In some embodiments, each R^(3b) is —C(═O)N(CH₃)₂.

In some embodiments, R⁴ is C₁₋₄ alkyl, halo, or cyano.

In some embodiments, R⁴ is C₁₋₄ alkyl.

In some embodiments, R⁴ is methyl or fluoro.

In some embodiments, R⁴ is methyl.

In some embodiments, R⁴ is halo.

In some embodiments, R⁴ is fluoro.

In some embodiments, R⁵ is halo, CN, or methyl.

In some embodiments, R⁵ is halo.

In some embodiments, R⁵ is chloro.

In some embodiments, Ar is:

In some embodiments, Ar is:

In some embodiments: Ar is:

R¹ is methyl;

R² is C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, or phenyl; wherein said phenyl isoptionally substituted by 1, 2, or 3 substituents independently selectedfrom halo, OH, CN, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkoxy;

R³ is Cy or C(═O)NR^(c)R^(d);

provided that either (i) R² is phenyl, wherein said phenyl is optionallysubstituted by 1, 2, or 3 substituents independently selected from halo,OH, CN, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkoxy; or (ii) R³ is Cy;

R⁴ is halo, C₁₋₄ alkyl, or C₁₋₄ haloalkyl;

R⁵ is halo, CN, C₁₋₄ alkyl, or C₁₋₄ haloalkyl;

R⁶ is H;

each Cy is independently selected from 4-6 membered heterocycloalkyl or5-6 membered heteroaryl, each of which is optionally substituted with 1or 2 independently selected R^(3b) groups;

each R^(c) and R^(d) is independently selected from H and C₁₋₆ alkyl;and

each R^(3b) is independently selected from halo, CN, C₁₋₆ alkyl, C₁₋₆haloalkyl, hydroxy, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, amino, C₁₋₆alkylamino, di(C₁₋₆ alkyl)amino, C₁₋₆ alkylsulfonyl, carbamyl, C₁₋₆alkylcarbamyl, and di(C₁₋₆ alkyl)carbamyl.

In some embodiments:

Ar is:

R² is C₁₋₆ alkoxy or phenyl, wherein said phenyl is optionallysubstituted by 1, 2, 3, or 4 independently selected halo groups;

R³ is C(═O)NR^(c)R^(d), 4-6 membered heterocycloalkyl, or 5-6 memberedheteroaryl; wherein said 4-6 membered heterocycloalkyl and 5-6 memberedheteroaryl is optionally substituted by 1, 2, or 3 independentlyselected R^(3b) groups;

each R^(3b) is independently selected from halo, CN, C₁₋₆ alkyl, C₁₋₆haloalkyl, hydroxy, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, amino, C₁₋₆alkylamino, di(C₁₋₆ alkyl)amino, C₁₋₆ alkylsulfonyl, carbamyl, C₁₋₆alkylcarbamyl, and di(C₁₋₆ alkyl)carbamyl;

R⁴ is C₁₋₄ alkyl or halo;

R⁵ is halo; and

R⁶ is H.

In some embodiments:

Ar is:

R² is C₁₋₆ alkoxy;

R³ is 4-6 membered heterocycloalkyl;

R⁴ and R⁵ are each independently halo; and

R⁶ is H.

In some embodiments:

Ar is:

R² is C₁₋₆ alkoxy or phenyl, wherein said phenyl is optionallysubstituted by 1, 2, 3, or 4 independently selected halo groups;

R³ is C(═O)NR^(c)R^(d), 4-6 membered heterocycloalkyl, or 5-6 memberedheteroaryl; wherein said 4-6 membered heterocycloalkyl and 5-6 memberedheteroaryl is optionally substituted by 1, 2, or 3 independentlyselected R^(3b) groups;

each R^(3b) is di(C₁₋₆ alkyl)carbamyl;

R⁴ is halo or C₁₋₄ alkyl;

R⁵ is halo; and

R⁶ is H.

In some embodiments, the compound is a compound having Formula (II):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is a compound having Formula (III):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is a compound having Formula (IIa):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is a compound having Formula (IIIa):

or a pharmaceutically acceptable salt thereof.

In some embodiments, R¹¹ is H.

In some embodiments, R¹² is H.

In some embodiments, R¹¹ and R¹² are each H.

In some embodiments, R¹³ is H.

In some embodiments, R¹⁴ is H.

In some embodiments, R¹⁵ is H.

In some embodiments, R¹¹, R¹³, R¹⁴ and R¹⁵ are each H.

In some embodiments, the compound is a compound selected from:

4-{3-chloro-6-ethoxy-2-fluoro-5-[1-([1,3]thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl]phenyl}pyrrolidin-2-one;

4-{3-chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-one;

5-{3-chloro-6-methoxy-2-methyl-5-[(1S)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}-N,N-dimethylpyridine-2-carboxamide;

4-chloro-N-ethyl-3′,5′-difluoro-3-methyl-6-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]biphenyl-2-carboxamide;

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is(S)-4-(3-chloro-6-ethoxy-2-fluoro-5-((S)-1-(thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl)phenyl)pyrrolidin-2-one;or a pharmaceutically acceptable salt thereof

In some embodiments, the compound is(R)-4-(3-chloro-6-ethoxy-2-fluoro-5-((R)-1-(thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl)phenyl)pyrrolidin-2-one;or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is(S)-4-(3-chloro-6-ethoxy-2-fluoro-5-((R)-1-(thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl)phenyl)pyrrolidin-2-one;or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is(R)-4-(3-chloro-6-ethoxy-2-fluoro-5-((S)-1-(thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl)phenyl)pyrrolidin-2-one;or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is(S)-4-(3-chloro-6-ethoxy-2-fluoro-5-((S)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl)phenyl)pyrrolidin-2-one;or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is(R)-4-(3-chloro-6-ethoxy-2-fluoro-5-((R)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl)phenyl)pyrrolidin-2-one;or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is(S)-4-(3-chloro-6-ethoxy-2-fluoro-5-((R)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl)phenyl)pyrrolidin-2-one;or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is(R)-4-(3-chloro-6-ethoxy-2-fluoro-5-((S)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl)phenyl)pyrrolidin-2-one;or a pharmaceutically acceptable salt thereof.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, can also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention which are, for brevity, described in thecontext of a single embodiment, can also be provided separately or inany suitable subcombination.

At various places in the present specification, divalent linkingsubstituents are described. It is specifically intended that eachdivalent linking substituent include both the forward and backward formsof the linking substituent. For example, —NR(CR′R″)_(n)— includes both—NR(CR′R″)_(n)— and —(CR′R″)_(n)NR—. Where the structure clearlyrequires a linking group, the Markush variables listed for that groupare understood to be linking groups.

The term “n-membered” where n is an integer typically describes thenumber of ring-forming atoms in a moiety where the number ofring-forming atoms is n. For example, piperidinyl is an example of a6-membered heterocycloalkyl ring, pyrazolyl is an example of a5-membered heteroaryl ring, pyridyl is an example of a 6-memberedheteroaryl ring, and 1,2,3,4-tetrahydro-naphthalene is an example of a10-membered cycloalkyl group.

As used herein, the phrase “optionally substituted” means unsubstitutedor substituted. As used herein, the term “substituted” means that ahydrogen atom is removed and replaced by a substituent. It is to beunderstood that substitution at a given atom is limited by valency.

Throughout the definitions, the term “C_(n-m)” indicates a range whichincludes the endpoints, wherein n and m are integers and indicate thenumber of carbons. Examples include C₁₋₄, C₁₋₆, and the like.

As used herein, the term “C_(n-m) alkyl”, employed alone or incombination with other terms, refers to a saturated hydrocarbon groupthat may be straight-chain or branched, having n to m carbons. In someembodiments, the alkyl group contains from 1 to 6 carbon atoms, from 1to 4 carbon atoms, from 1 to 3 carbon atoms, or 1 to 2 carbon atoms.Examples of alkyl moieties include, but are not limited to, chemicalgroups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl,isobutyl, sec-butyl; higher homologs such as 2-methyl-1-butyl, n-pentyl,3-pentyl, n-hexyl, 1,2,2-trimethylpropyl, and the like.

As used herein, “C_(n-m) alkenyl” refers to an alkyl group having one ormore double carbon-carbon bonds and having n to m carbons. In someembodiments, the alkenyl moiety contains 2 to 6, 2 to 4, or 2 to 3carbon atoms. Example alkenyl groups include, but are not limited to,ethenyl, n-propenyl, isopropenyl, n-butenyl, sec-butenyl, and the like.

As used herein, “C_(n-m) alkynyl” refers to an alkyl group having one ormore triple carbon-carbon bonds and having n to m carbons. Examplealkynyl groups include, but are not limited to, ethynyl, propyn-1-yl,propyn-2-yl, and the like. In some embodiments, the alkynyl moietycontains 2 to 6, 2 to 4, or 2 to 3 carbon atoms.

As used herein, the term “alkylene”, employed alone or in combinationwith other terms, refers to a divalent alkyl linking group. Examples ofalkylene groups include, but are not limited to, ethan-1,2-diyl,propan-1,3-diyl, propan-1,2-diyl, butan-1,4-diyl, butan-1,3-diyl,butan-1,2-diyl, 2-methyl-propan-1,3-diyl, and the like.

As used herein, the term “C_(n-m) alkoxy”, employed alone or incombination with other terms, refers to a group of formula —O-alkyl,wherein the alkyl group has n to m carbons. Example alkoxy groupsinclude methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy),t-butoxy, and the like. In some embodiments, the alkyl group has 1 to 6,1 to 4, or 1 to 3 carbon atoms.

As used herein, the term “C_(n-m) alkylamino” refers to a group offormula —NH(alkyl), wherein the alkyl group has n to m carbon atoms. Insome embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbonatoms.

As used herein, the term “C_(n-m) alkoxycarbonyl” refers to a group offormula —C(O)O-alkyl, wherein the alkyl group has n to m carbon atoms.In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3carbon atoms.

As used herein, the term “C_(n-m) alkylcarbonyl” refers to a group offormula —C(O)-alkyl, wherein the alkyl group has n to m carbon atoms. Insome embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbonatoms.

As used herein, the term “C_(n-m) alkylcarbonylamino” refers to a groupof formula —NHC(O)-alkyl, wherein the alkyl group has n to m carbonatoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to3 carbon atoms.

As used herein, the term “C_(n-m) alkylsulfonylamino” refers to a groupof formula —NHS(O)₂-alkyl, wherein the alkyl group has n to m carbonatoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to3 carbon atoms.

As used herein, the term “aminosulfonyl” refers to a group of formula—S(O)₂NH₂.

As used herein, the term “C_(n-m) alkylaminosulfonyl” refers to a groupof formula —S(O)₂NH(alkyl), wherein the alkyl group has n to m carbonatoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to3 carbon atoms.

As used herein, the term “di(C_(n-m) alkyl)aminosulfonyl” refers to agroup of formula —S(O)₂N(alkyl)₂, wherein each alkyl group independentlyhas n to m carbon atoms. In some embodiments, each alkyl group has,independently, 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

As used herein, the term “aminosulfonylamino” refers to a group offormula —NHS(O)₂NH₂.

As used herein, the term “C_(n-m) alkylaminosulfonylamino” refers to agroup of formula —NHS(O)₂NH(alkyl), wherein the alkyl group has n to mcarbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4,or 1 to 3 carbon atoms.

As used herein, the term “di(C_(n-m) alkyl)aminosulfonylamino” refers toa group of formula —NHS(O)₂N(alkyl)₂, wherein each alkyl groupindependently has n to m carbon atoms. In some embodiments, each alkylgroup has, independently, 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

As used herein, the term “aminocarbonylamino”, employed alone or incombination with other terms, refers to a group of formula —NHC(O)NH₂.

As used herein, the term “C_(n-m) alkylaminocarbonylamino” refers to agroup of formula —NHC(O)NH(alkyl), wherein the alkyl group has n to mcarbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4,or 1 to 3 carbon atoms.

As used herein, the term“di(C_(n-m alkyl)aminocarbonylamino” refers to a group of formula —NHC(O)N(alkyl))₂, wherein each alkyl group independently has n to m carbon atoms. Insome embodiments, each alkyl group has, independently, 1 to 6, 1 to 4,or 1 to 3 carbon atoms.

As used herein, the term “C_(n-m) alkylcarbamyl” refers to a group offormula —C(O)—NH(alkyl), wherein the alkyl group has n to m carbonatoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to3 carbon atoms.

As used herein, the term “thio” refers to a group of formula —SH.

As used herein, the term “C_(n-m) alkylthio” refers to a group offormula —S-alkyl, wherein the alkyl group has n to m carbon atoms. Insome embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbonatoms.

As used herein, the term “C_(n-m) alkylsulfinyl” refers to a group offormula —S(O)-alkyl, wherein the alkyl group has n to m carbon atoms. Insome embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbonatoms.

As used herein, the term “C_(n-m) alkylsulfonyl” refers to a group offormula —S(O)₂-alkyl, wherein the alkyl group has n to m carbon atoms.In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3carbon atoms.

As used herein, the term “amino” refers to a group of formula —NH₂.

As used herein, the term “carbamyl” to a group of formula —C(O)NH₂.

As used herein, the term “carbonyl”, employed alone or in combinationwith other terms, refers to a —C(═O)— group. Also may be written asC(O).

As used herein, the term “cyano-C₁₋₃ alkyl” refers to a group of formula—(C₁₋₃ alkylene)-CN.

As used herein, the term “HO—C₁₋₃ alkyl” refers to a group of formula—(C₁₋₃ alkylene)-OH.

As used herein, the term “C₁₋₃ alkoxy-C₁₋₃ alkyl” refers to a group offormula —(C₁₋₃ alkylene)-O(C₁₋₃ alkyl).

As used herein, the term “C₁₋₄ alkoxy-C₁₋₄ alkyl” refers to a group offormula —(C₁₋₄ alkylene)-O(C₁₋₄ alkyl).

As used herein, the term “C₁₋₄ haloalkoxy-C₁₋₄ alkyl” refers to a groupof formula —(C₁₋₄ alkylene)-O(C₁₋₄ haloalkyl).

As used herein, the term “carboxy” refers to a group of formula —C(O)OH.

As used herein, the term “di(C_(n-m)-alkyl)amino” refers to a group offormula —N(alkyl)₂, wherein the two alkyl groups each has,independently, n to m carbon atoms. In some embodiments, each alkylgroup independently has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

As used herein, the term “di(C_(n-m)-alkyl)carbamyl” refers to a groupof formula —C(O)N(alkyl)₂, wherein the two alkyl groups each has,independently, n to m carbon atoms. In some embodiments, each alkylgroup independently has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

As used herein, “halo” refers to F, Cl, Br, or I. In some embodiments,the halo group is F or Cl.

As used herein, “C_(n-m) haloalkoxy” refers to a group of formula—O-haloalkyl having n to m carbon atoms. An example haloalkoxy group isOCF₃. In some embodiments, the haloalkoxy group is fluorinated only. Insome embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbonatoms.

As used herein, the term “C_(n-m) haloalkyl”, employed alone or incombination with other terms, refers to an alkyl group having from onehalogen atom to 2s+1 halogen atoms which may be the same or different,where “s” is the number of carbon atoms in the alkyl group, wherein thealkyl group has n to m carbon atoms. In some embodiments, the haloalkylgroup is fluorinated only. In some embodiments, the alkyl group has 1 to6, 1 to 4, or 1 to 3 carbon atoms.

As used herein, “cycloalkyl” refers to non-aromatic cyclic hydrocarbonsincluding cyclized alkyl and/or alkenyl groups. Cycloalkyl groups caninclude mono- or polycyclic (e.g., having 2, 3 or 4 fused rings) groupsand spirocycles. Cycloalkyl groups can have 3, 4, 5, 6, or 7ring-forming carbons (C₃₋₇). Ring-forming carbon atoms of a cycloalkylgroup can be optionally substituted by oxo or sulfido (e.g., C(O) orC(S)). Cycloalkyl groups also include cycloalkylidenes. Examplecycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl,cycloheptatrienyl, norbornyl, norpinyl, norcarnyl, and the like. In someembodiments, cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl. Also included in the definition of cycloalkyl are moietiesthat have one or more aromatic rings fused (i.e., having a bond incommon with) to the cycloalkyl ring, for example, benzo or thienylderivatives of cyclopentane, cyclohexane, and the like. A cycloalkylgroup containing a fused aromatic ring can be attached through anyring-forming atom including a ring-forming atom of the fused aromaticring.

As used herein, “heteroaryl” refers to a monocyclic or polycyclicaromatic heterocycle having at least one heteroatom ring member selectedfrom sulfur, oxygen, and nitrogen. In some embodiments, the heteroarylring has 1, 2, 3, or 4 heteroatom ring members independently selectedfrom nitrogen, sulfur and oxygen. In some embodiments, any ring-formingN in a heteroaryl moiety can be an N-oxide. In some embodiments, theheteroaryl has 5-10 ring atoms and 1, 2, 3 or 4 heteroatom ring membersindependently selected from nitrogen, sulfur and oxygen. In someembodiments, the heteroaryl has 5-6 ring atoms and 1 or 2 heteroatomring members independently selected from nitrogen, sulfur and oxygen. Insome embodiments, the heteroaryl is a five-membered or six-memberetedheteroaryl ring.

A five-membered heteroaryl ring is a heteroaryl with a ring having fivering atoms wherein one or more (e.g., 1, 2, or 3) ring atoms areindependently selected from N, O, and S. Exemplary five-membered ringheteroaryls are thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl,oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl, 1,2,3-triazolyl,tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-triazolyl,1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl,1,3,4-thiadiazolyl, and 1,3,4-oxadiazolyl.

A six-membered heteroaryl ring is a heteroaryl with a ring having sixring atoms wherein one or more (e.g., 1, 2, or 3) ring atoms areindependently selected from N, O, and S. Exemplary six-membered ringheteroaryls are pyridyl, pyrazinyl, pyrimidinyl, triazinyl andpyridazinyl.

As used herein, “heterocycloalkyl” refers to non-aromatic monocyclic orpolycyclic heterocycles having one or more ring-forming heteroatomsselected from O, N, or S. Included in heterocycloalkyl are monocyclic4-, 5-, 6-, and 7-membered heterocycloalkyl groups. Heterocycloalkylgroups can also include spirocycles. Example heterocycloalkyl groupsinclude pyrrolidin-2-one, 1,3-isoxazolidin-2-one, pyranyl,tetrahydropuran, oxetanyl, azetidinyl, morpholino, thiomorpholino,piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl,pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl,oxazolidinyl, thiazolidinyl, imidazolidinyl, azepanyl, benzazapene, andthe like. Ring-forming carbon atoms and heteroatoms of aheterocycloalkyl group can be optionally substituted by oxo or sulfido(e.g., C(O), S(O), C(S), or S(O)₂, etc.). The heterocycloalkyl group canbe attached through a ring-forming carbon atom or a ring-formingheteroatom. In some embodiments, the heterocycloalkyl group contains 0to 3 double bonds. In some embodiments, the heterocycloalkyl groupcontains 0 to 2 double bonds. Also included in the definition ofheterocycloalkyl are moieties that have one or more aromatic rings fused(i.e., having a bond in common with) to the cycloalkyl ring, forexample, benzo or thienyl derivatives of piperidine, morpholine,azepine, etc. A heterocycloalkyl group containing a fused aromatic ringcan be attached through any ring-forming atom including a ring-formingatom of the fused aromatic ring. In some embodiments, theheterocycloalkyl has 4-10, 4-7 or 4-6 ring atoms with 1 or 2 heteroatomsindependently selected from nitrogen, oxygen, or sulfur and having oneor more oxidized ring members.

As used herein, the term “phenyl-C_(n-m)-alkyl”, employed alone or incombination with other terms, refers to a group of formula-alkylene-phenyl, wherein the alkylene portion has n to m carbon atoms.In some embodiments, the alkylene portion has 1 to 3, 1 to 2, or 1carbon atom(s). In some embodiments, the alkylene portion is methylene.In some embodiments, the arylalkyl group is benzyl.

As used herein, the term “C_(n-m) heteroaryl-C_(o-p)-alkyl”, employedalone or in combination with other terms, refers to a group of formula-alkylene-heteroaryl, wherein the heteroaryl portion has n to m ringmember carbon atoms and the alkylene portion has o to p carbon atoms. Insome embodiments, the alkylene portion has 1 to 3, 1 to 2, or 1 carbonatom(s). In some embodiments, the alkylene portion is methylene.

As used herein, the term “heterocycloalkyloxy” refers to a group offormula —O-(heterocycloalkyl).

At certain places, the definitions or embodiments refer to specificrings (e.g., an azetidine ring, a pyridine ring, etc.). Unless otherwiseindicated, these rings can be attached to any ring member provided thatthe valency of the atom is not exceeded. For example, an azetidine ringmay be attached at any position of the ring, whereas an azetidin-3-ylring is attached at the 3-position.

The compounds described herein can be asymmetric (e.g., having one ormore stereocenters). All stereoisomers, such as enantiomers anddiastereomers, are intended unless otherwise indicated. Compounds of thepresent application that contain asymmetrically substituted carbon atomscan be isolated in optically active or racemic forms. Methods on how toprepare optically active forms from optically inactive startingmaterials are known in the art, such as by resolution of racemicmixtures or by stereoselective synthesis. Many geometric isomers ofolefins, C═N double bonds, and the like can also be present in thecompounds described herein, and all such stable isomers are contemplatedin the present application. Cis and trans geometric isomers of thecompounds of the present application are described and may be isolatedas a mixture of isomers or as separated isomeric forms.

In some embodiments, the compound has the (R)-configuration (e.g., atthe carbon to which R¹ is attached). In some embodiments, the compoundhas the (S)-configuration (e.g., at the carbon to which R¹ is attached).

Resolution of racemic mixtures of compounds can be carried out by any ofnumerous methods known in the art. An example method includes fractionalrecrystallizaion using a chiral resolving acid which is an opticallyactive, salt-forming organic acid. Suitable resolving agents forfractional recrystallization methods are, for example, optically activeacids, such as the D and L forms of tartaric acid, diacetyltartaricacid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid orthe various optically active camphorsulfonic acids such asβ-camphorsulfonic acid. Other resolving agents suitable for fractionalcrystallization methods include stereoisomerically pure forms ofα-methylbenzylamine (e.g., S and R forms, or diastereomerically pureforms), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine,cyclohexylethylamine, 1,2-diaminocyclohexane, and the like.

Resolution of racemic mixtures can also be carried out by elution on acolumn packed with an optically active resolving agent (e.g.,dinitrobenzoylphenylglycine). Suitable elution solvent composition canbe determined by one skilled in the art.

Compounds of the invention also include tautomeric forms. Tautomericforms result from the swapping of a single bond with an adjacent doublebond together with the concomitant migration of a proton. Tautomericforms include prototropic tautomers which are isomeric protonationstates having the same empirical formula and total charge. Exampleprototropic tautomers include ketone-enol pairs, amide-imidic acidpairs, lactam-lactim pairs, enamine-imine pairs, and annular forms wherea proton can occupy two or more positions of a heterocyclic system, forexample, 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and2H-isoindole, and 1H- and 2H-pyrazole. Tautomeric forms can be inequilibrium or sterically locked into one form by appropriatesubstitution.

Compounds of the invention can also include all isotopes of atomsoccurring in the intermediates or final compounds. Isotopes includethose atoms having the same atomic number but different mass numbers.For example, isotopes of hydrogen include tritium and deuterium.

The term, “compound,” as used herein is meant to include allstereoisomers, geometric isomers, tautomers, and isotopes of thestructures depicted. Compounds herein identified by name or structure asone particular tautomeric form are intended to include other tautomericforms unless otherwise specified.

All compounds, and pharmaceutically acceptable salts thereof, can befound together with other substances such as water and solvents (e.g.hydrates and solvates) or can be isolated.

In some embodiments, the compounds of the invention, or salts thereof,are substantially isolated. By “substantially isolated” is meant thatthe compound is at least partially or substantially separated from theenvironment in which it was formed or detected. Partial separation caninclude, for example, a composition enriched in the compounds of theinvention. Substantial separation can include compositions containing atleast about 50%, at least about 60%, at least about 70%, at least about80%, at least about 90%, at least about 95%, at least about 97%, or atleast about 99% by weight of the compounds of the invention, or saltthereof. Methods for isolating compounds and their salts are routine inthe art.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The expressions, “ambient temperature” and “room temperature” or “rt” asused herein, are understood in the art, and refer generally to atemperature, e.g. a reaction temperature, that is about the temperatureof the room in which the reaction is carried out, for example, atemperature from about 20° C. to about 30° C.

The present application also includes pharmaceutically acceptable saltsof the compounds described herein. As used herein, “pharmaceuticallyacceptable salts” refers to derivatives of the disclosed compoundswherein the parent compound is modified by converting an existing acidor base moiety to its salt form. Examples of pharmaceutically acceptablesalts include, but are not limited to, mineral or organic acid salts ofbasic residues such as amines; alkali or organic salts of acidicresidues such as carboxylic acids; and the like. The pharmaceuticallyacceptable salts of the present application include the conventionalnon-toxic salts of the parent compound formed, for example, fromnon-toxic inorganic or organic acids. The pharmaceutically acceptablesalts of the present application can be synthesized from the parentcompound which contains a basic or acidic moiety by conventionalchemical methods. Generally, such salts can be prepared by reacting thefree acid or base forms of these compounds with a stoichiometric amountof the appropriate base or acid in water or in an organic solvent, or ina mixture of the two; generally, non-aqueous media like ether, ethylacetate, alcohols (e.g., methanol, ethanol, iso-propanol, or butanol) oracetonitrile (ACN) are preferred. Lists of suitable salts are found inRemington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company,Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2(1977), each of which is incorporated herein by reference in itsentirety.

Synthesis

Compounds described herein, including salts thereof, can be preparedusing known organic synthesis techniques and can be synthesizedaccording to any of numerous possible synthetic routes. In someembodiments, the compounds can be prepared as described in U.S. patentapplication Ser. No. 13/601,349, filed Aug. 31, 2012, which isincorporated herein by reference in its entirety.

The reactions for preparing compounds described herein can be carriedout in suitable solvents which can be readily selected by one of skillin the art of organic synthesis. Suitable solvents can be substantiallynon-reactive with the starting materials (reactants), the intermediates,or products at the temperatures at which the reactions are carried out,e.g., temperatures which can range from the solvent's freezingtemperature to the solvent's boiling temperature. A given reaction canbe carried out in one solvent or a mixture of more than one solvent.Depending on the particular reaction step, suitable solvents for aparticular reaction step can be selected by the skilled artisan.

Preparation of compounds described herein can involve the protection anddeprotection of various chemical groups. The need for protection anddeprotection, and the selection of appropriate protecting groups, can bereadily determined by one skilled in the art. The chemistry ofprotecting groups can be found, for example, in T. W. Greene and P. G.M. Wuts, Protective Groups in Organic Synthesis, 3^(rd) Ed., Wiley &Sons, Inc., New York (1999), which is incorporated herein by referencein its entirety.

Reactions can be monitored according to any suitable method known in theart. For example, product formation can be monitored by spectroscopicmeans, such as nuclear magnetic resonance spectroscopy (e.g., ¹H or¹³C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), massspectrometry, or by chromatographic methods such as high performanceliquid chromatography (HPLC), liquid chromatography-mass spectroscopy(LCMS), or thin layer chromatography (TLC). Compounds can be purified bythose skilled in the art by a variety of methods, including highperformance liquid chromatography (HPLC) (“Preparative LC-MSPurification: Improved Compound Specific Method Optimization” Karl F.Blom, Brian Glass, Richard Sparks, Andrew P. Combs J. Combi. Chem. 2004,6(6), 874-883, which is incorporated herein by reference in itsentirety) and normal phase silica chromatography.

For example, compounds of Formula I can be formed as shown in Scheme I.Compound (i) can be halogenated with N-chlorosuccinamide,N-bromosuccinamide or N-iodosuccinamide to give compound (ii) whereX═Cl, Br, or I. The halo group of (ii) can be coupled to R³-M, where Mis a boronic acid, boronic ester or an appropriately substituted metal(e.g., R³-M is R³—B(OH)₂ or R³—Sn(Bu)₄), under standard Suzukiconditions or standard Stille conditions (e.g., in the presence of apalladium(0) catalyst, such as tetrakis(triphenylphosphine)palladium(0)and a base (e.g., a bicarbonate or carbonate base) to give a derivativeof formula (iii). Alternatively, R³-M can be a cyclic amine (where M isH and attached to the amine nitrogen) with coupling to compound (ii)being performed by heating in base or under Buchwald conditions (e.g.,in the presence of a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0) and a base (e.g., an alkoxidebase)) to afford ketone (iii). Reductive amination of ketone (iii) canfurnish the amine intermediate (v). Alternatively, ketone (iii) can bereduced to give an alcohol which can be converted to the mesylate andreacted with sodium azide to give an azide derivative (iv). The azide ofcompound (iv) can be converted to an amine (v) under appropriatereducing conditions, such as trimethylphosphine or TMSI. Finally thesecondary amine can be reacted with a heteroaryl halide compound (e.g.,Ar—X, such as 7-chloro[1,3]thiazolo[5,4-d]pyrimidine or4-chloropyrido[3,2-d]pyrimidine) to give a compound of Formula I (vi).

Alternatively, compounds of Formula I can also be formed as shown inScheme II. Ketone (i) can be halogenated with N-chlorosuccinamide,N-bromosuccinamide or N-iodosuccinamide to give compound (ii) whereX═Cl, Br, or I. Ketone (ii) can be reduced to give an alcohol (iii)which can be converted to the mesylate and reacted with sodium azide togive an azide derivative (iv). The azide of compound (iv) can beconverted to an amine (v) under appropriate reducing conditions, such astrimethylphosphine or TMSI. The amine (v) can be protected with asuitable protecting group (e.g., by reacting with Boc₂O) and purified bychiral chromatography to afford a single enantiomer of amine compound(v). The amino group can be deprotected (e.g., TFA when P=Boc) and theresulting amine can be reacted with a heteroaryl halide compound (e.g.,Ar—X) to give compound (vi). Finally, the halo group of (vi) can becoupled to R³-M, where M is a boronic acid, boronic ester or anappropriately substituted metal (e.g., R³-M is R³—B(OH)₂ or R³—Sn(Bu)₄),under standard Suzuki conditions or standard Stille conditions (e.g., inthe presence of a palladium(0) catalyst, such astetrakis(triphenylphosphine)-palladium(0) and a base (e.g., abicarbonate or carbonate base)) to give a derivative of Formula I (vii).Alternatively, R³-M can be a cyclic amine (where M is H and attached tothe amine nitrogen) with coupling to compound (vi) being performed byheating in base or under Buchwald conditions (e.g., in the presence of apalladium(0) catalyst, such as tetrakis(triphenylphosphine)-palladium(0)and a base (e.g., an alkoxide base)) to afford compounds of Formula I(vii).

Compounds of Formula I wherein L is O can be formed as shown in SchemeIII. The phenols (i) can be alkylated using Mitsunobu conditions (e.g.,ROH, DEAD, Ph₃P) or standard alkylating conditions (R-Lg, Lg=leavinggroup) to afford ether derivatives (ii). The halo group (e.g., X═Br, I)of (ii) can be coupled to R³-M, where M is a boronic acid, boronicester, or an appropriately substituted metal (e.g., R³-M is R³—B(OH)₂ orR³—Sn(Bu)₄), under standard Suzuki conditions or standard Stilleconditions (e.g., in the presence of a palladium(0) catalyst, such astetrakis(triphenylphosphine)-palladium(0) and a base (e.g., abicarbonate or carbonate base)) to give a derivative of formula (iii).Alternatively, R³-M can be a cyclic amine (where M is H and attached tothe amine nitrogen) with coupling to compound (ii) being performed byheating in base or under Buchwald conditions (e.g., in the presence of apalladium(0) catalyst, such as tetrakis(triphenylphosphine)-palladium(0)and a base (e.g., an alkoxide base)) to afford compounds of formula(iii). The ketone (iii) can be transformed using similar methods asshown in Schemes I and II to afford compounds of Formula I (iv).Alternatively, the halo-ketone (ii) can be transformed using similarmethods as shown in Schemes I and II to afford halo intermediate (v).Suzuki, Stille, or Buchwald coupling of R³-M with halo intermediate (v)by similar methods described in Schemes I and II can afford compounds ofFormula I (iv).

Compounds of Formula I can be formed as shown in Scheme IV. Compound (i)can be acylated with a suitable acylating reagent (e.g., R¹—COCl) toform an ester which can be rearranged under Lewis acid conditions (e.g.,BF₃/HOAc complex) to afford ketone (ii). Halogenation of ketone (ii)using NXS (e.g., NXS=N-chlorosuccinamide, N-bromosuccinamide orN-iodosuccinamide) can give compound (iii) where X═Cl, Br, or I. Thephenol can be converted to the triflate (iv) using standard conditions(e.g.,Tf₂O). The triflate group of (iv) can be coupled to R²-M, where Mis a boronic acid, boronic ester or an appropriately substituted metal(e.g., R²-M is R²—B(OH)₂ or R²—Sn(Bu)₄), under standard Suzukiconditions or standard Stille conditions (e.g., in the presence of apalladium(0) catalyst, such as tetrakis(triphenylphosphine)palladium(0)and a base (e.g., a bicarbonate or carbonate base)) to give ketone (v).Alternatively, R²-M can be a cyclic amine (where M is H and attached tothe amine nitrogen) with coupling to compound (iv) being performed byheating in base or under Buchwald conditions (e.g., in the presence of apalladium(0) catalyst, such as tetrakis(triphenylphosphine)palladium(0)and a base (e.g., an alkoxide base)) to afford ketone (v) . The halogroup of (v) can be coupled to R³-M, where M is a boronic acid, boronicester or an appropriately substituted metal (e.g., R³-M is R³—B(OH)₂ orR³—Sn(Bu)₄), under standard Suzuki conditions or standard Stilleconditions (e.g., in the presence of a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0) and a base (e.g., a bicarbonateor carbonate base)) to give a derivative of formula (vi). Alternatively,R³-M can be a cyclic amine (where M is H and attached to the aminenitrogen) with coupling to compound (v) being performed by heating inbase or under Buchwald conditions (e.g., in the presence of apalladium(0) catalyst, such as tetrakis(triphenylphosphine)palladium(0)and a base (e.g., an alkoxide base)) to afford ketone (vi). Ketone (vi)can be transformed using similar methods as shown in Schemes I and II toafford compounds of Formula I (vii).

Alternatively, the halo-ketone (v) can be transformed using similarmethods as shown in Schemes I and II to afford halo intermediate (viii).Suzuki, Stille, or Buchwald coupling of M-R³ with compound (viii) bysimilar methods described in Schemes I and II can also afford compoundsof Formula I (vii).

Ketones which can be used in the processes of Schemes I-III can beformed as shown in Scheme V. The carboxylic acid (i) can be activatedwith a coupling agent (e.g., HBTU, HATU or EDC) and then reacted withN,O-dimethylhydroxylamine to give a N-methoxy-N-methylcarboxamidederivative (ii). Amide (ii) may then be reacted with a Grignard reagentof formula R¹—MgX (X=halo) to give ketone (iii). Ketone (iii) can betransformed using similar methods as shown in Schemes I, II and III toafford compounds of Formula I.

Compounds which can be used in the processes of Schemes I-III can alsobe formed as shown in Scheme VI. The halo-ketone (i) can be converted tothe cyano-ketone (ii) using standard cyanation conditions (e.g., Pd(0)and Zn(CN)₂). Hydrolysis of the cyano group of (ii) under acidic orbasic conditions can give the carboxylic acid which can be coupled toamines using a coupling agent (e.g., HATU, HBTU, EDC) and appropriateamines (HNR^(c)R^(d)) to give amide (iii) (R^(c) and R^(d) are variousoptionally substituted cyclic groups and non-cyclic groups, or R^(c) andR^(d), along with the nitrogen atom to which they are attached cancyclize to form a heterocycloalkyl group). The ketone of amide (iii) canbe transformed using similar methods as shown in Schemes I, II and IIIto afford compounds of Formula I (v).

Additional compounds which can be used in the processes of Schemes I-IIIcan be formed as shown in Scheme VII. Ketone (i) can be converted to thenitro-ketone (ii) using standard nitration conditions (e.g., HNO₃).Reduction of the nitro group of (ii) under standard conditions (e.g.,Fe, Zn, or H₂ over Pd/C) can give the amino compound which can bederivatized, including acylated with appropriate acylating agents (e.g.,R^(b)C(═O)Cl, R^(a)OC(═O)Cl, and (R^(c)R^(d))NC(═O)Cl) to give ketone(iii) R^(a), R^(b), R^(c), and R^(d), for example, can be variousoptionally substituted cyclic groups and non-cyclic groups as defined inthe claims and throughout). Ketone (iii) can be transformed usingsimilar methods as shown in Schemes I, II and III to afford compounds ofFormula I (v).

Further compounds which can be used in the processes of Schemes I-IIIcan be formed as shown in Scheme VIII. Ether (i) can be converted to aphenol (ii) using standard conditions (e.g., BBr₃). The halo-phenol (ii)can be converted to the cyano-phenol (iii) using standard cyanationconditions (e.g., CuCN or Pd(0) and Zn(CN)₂). The phenol (iii) can beconverted to the triflate (iv) using Tf₂O. The triflate group of (iv)can be coupled to R²-M, where M is a boronic acid, boronic ester, or anappropriately substituted metal (e.g., R²-M is R²—B(OH)₂ or R²—Sn(Bu)₄),under standard Suzuki conditions or standard Stille conditions (e.g., inthe presence of a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0) and a base (e.g., a bicarbonateor carbonate base)) to give a derivative of formula (v). Alternatively,R²-M can be a cyclic amine (where M is H and attached to the aminenitrogen) with coupling to compound (iv) being performed by heating inbase or under Buchwald conditions (e.g., in the presence of apalladium(0) catalyst, such as tetrakis(triphenylphosphine)palladium(0)and a base (e.g., an alkoxide base)) to afford ketone (v). Hydrolysis ofthe cyano group of (v) under acidic or basic conditions can give thecarboxylic acid which can be coupled to amines using a coupling agent(e.g., HATU, HBTU, EDC) and an appropriate amine (HNR^(c)R^(d)) to giveamide (vi). The ketone group of amide (vi) can be transformed usingsimilar methods as shown in Schemes I, II and III to afford compounds ofFormula I (vii).

Ketones which can be used in the processes of Scheme I, II and III, canalso be formed as shown in Scheme IX. The carboxylic acid (i) can beactivated with a coupling agent (e.g. HBTU or HATU) and then reactedwith N,O-dimethylhydroxylamine to give a N-methoxy-N-methylcarboxamide.The phenols can be alkylated using Mitsunobu conditions (e.g., ROH,DEAD, Ph₃P) or standard alkylating conditions (R′-Lg, Lg=leaving group)to afford ether derivatives (ii). The halo group of (ii) (X¹ is halo)can be coupled to R³-M, where M is a boronic acid, boronic ester or anappropriately substituted metal (e.g., R³-M is R³—B(OH)₂, R³—Sn(Bu)₄, orZn—R³), under standard Suzuki conditions or standard Stille conditions(e.g., in the presence of a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0) and a base (e.g., a bicarbonateor carbonate base) or standard Negishi conditions (e.g., in the presenceof a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0), to give a derivative offormula (iii).

Alternatively, R³-M can be a cyclic amine (where M is H and attached tothe amine nitrogen) with coupling to compound (ii) being performed byheating in base or under Buchwald conditions (e.g., in the presence of apalladium(0) catalyst, such as tetrakis(triphenylphosphine)palladium(0)and a base (e.g., an alkoxide base)) to afford amides (iii). Reaction ofcompound (iii) with a Grignard reagent of formula R¹—MgX² (X²=halo) cangive ketone (iv). Ketone (iv) can be transformed using similar methodsas shown in Schemes I, II and III to afford compounds of Formula I.

Ketones which can be used in the processes of Schemes I, II and III, canalso be formed as shown in Scheme X below. The halo group (e.g., X¹ ═I)of (i) can be coupled to a zinc reagent R³—Zn (e.g., such as tert-butyl3-iodoazetidine-1-carboxylate with Zn dust) under standardKnochel/Negishi conditions (e.g., in the presence of a palladium(0)catalyst, such as tri-(2-furyl)phosphine andtris(dibenzylideneacetone)dipalladium(0) and 1,2-dibromoethane andchlorotrimethylsilane) to give ketone (ii). The azetidine (ii) can bedeprotected (e.g., Pg=Boc, using TFA) and then reacted under alkylating,acylating or reductive amination (e.g., RX such as R—Br, RC(═O)Cl,R—S(O₂)Cl, RN═C═O or RCHO and a reducing agent) conditions to affordketone derivatives (iii) which can be converted to compounds of FormulaI (v) by similar methods shown in Schemes I, II, and III. Alternatively,ketone (ii) can be converted to compounds of formula (vii) using similarmethods as shown in Schemes I, II and III. The protecting group on theamine of compound (vii) can be removed under standard conditions andthen reacted under alkylating, acylating or reductive aminationconditions (e.g., RX such as R—Br, RC(═O)Cl, R—S(O₂)Cl, RN═C═O or RCHOand a reducing agent) to give compounds of Formula I (v).

Compounds of Formula I can also be formed as shown in Scheme XI. Thehalo group of (i) can be coupled to an alkene (e.g., acrylate oracrylamide) under standard Heck conditions (e.g., in the presence of apalladium(II) catalyst, such as palladium acetate) to give an alkene offormula (ii). Reaction of alkene (ii) with nitromethane in the presenceof DBU can afford the nitro derivative (iii) which can be reduced understandard conditions (e.g., NiCl₂/NaBH₄ or Raney Ni) to give a free aminewhich cyclizes to form lactam (iv). The lactam can be alkylated understandard conditions (e.g., R^(3a)—X², where X²=halo, in the presence ofa base, such as TEA or NaH) to give an N-alkyl-lactam (v). Compounds offormula (v), and pyrrolidines derived from the reduction of the lactam(v) with suitable reducing agents, such as LiAlH₄, can be converted tocompounds of Formula I using conditions described in Schemes I, II andIII.

Compounds of Formula I can also be formed as shown in Scheme XII. Thehalo group of (i) can be coupled to R³-M, where M is an appropriatelysubstituted metal (e.g., R³-M is R³B(OH)₂; appropriate non-limitingstarting materials for generating R³-M are shown in Scheme XII) understandard Suzuki conditions (e.g., in the presence of a palladium(0)catalyst, such as tetrakis(triphenylphosphine)palladium(0)) to give analkene of formula (ii). Epoxidation of alkene (ii) with mCPBA can affordthe epoxide (iii) which can be reacted with a secondary or primary amine(amine=NHR^(c)R^(d); R^(c)═H for primary amine) to give amino compoundsof formula (iv). Secondary or tertiary amine derivatives (iv) can befurther reacted with carbonyldiamidazole (CDI) or phosgene to form anoxazolidinone (v) or an acetyl-halide (e.g., chloro-acetylchloride inthe presence of base, such as TEA) to give the N-acyl derivative whichcan be converted to the morpholinone derivative (vi) upon treatment witha base (e.g., NaH). Compounds of formula (iv, v, and vi) can bedeprotected using standard conditions (e.g., compounds protected withTHP groups may be treated with an acid, such as TFA or HCl) to givecompounds of Formula I (iv, v, and vi).

Compounds of Formula I can also be formed as shown in Scheme XIII.Sharpless amino-hydroxylation of an alkene of formula (i) under suitableconditions (A or B, as described in JACS, 2001, 123(9), 1862-1871 and J.Org. Chem., 2011, 76, 358-372) can give either amino-hydroxy isomer (ii)or (iii). Compound (ii) can be reacted with carbonyldiamidazole orphosgene to form an oxazolidinone (iv), or an acetyl-halide (e.g.,chloro-acetylchloride in the presence of base, such as TEA) to give anN-acyl derivative which can be converted to the morpholinone derivative(v) upon treatment with a base (e.g., NaH). The alternate amino-hydroxyisomer (iii) can be converted to oxazolidinone and morpholinonederivatives as shown in Scheme XII.

Compounds of Formula I can be synthesized as shown in Scheme XIV. Thehalo group (e.g., X¹═Cl, Br, I) of (i) can be converted to the boronateester (ii) under standard conditions (e.g., pinacol boronate ester inthe presence of a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0)). Boronate (ii) can be reactedwith an arylhalide or heteroarylhalide (e.g., R³—X²) under Suzukiconditions (e.g., in the presence of a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0) and a base, such as Na₂CO₃) togive formula (iii). Formula (iii) can be converted to Formula I usingthe reaction conditions described in Schemes I, II or III.

Compounds of Formula I, where R⁴═F or CN, can be formed as shown inScheme XV. Compound (i) can be acylated with a suitable acylatingreagent (e.g., R¹—COCl) to form an ester which can be rearranged underLewis acid conditions (e.g., BF₃/HOAc complex) to afford ketone (ii).Ketone (ii) can be halogenated with N-chlorosuccinamide,N-bromosuccinamide or N-iodosuccinamide to give phenol (iii), whereX¹═Cl, Br, or I. Compound (iii) can be alkylated (e.g. R²—X² and a base,such as NaH or Na₂CO₃; or under Mitsunobu conditions) to afford theether (iv). The fluoro group of (iv) can be displaced (e.g., with NaCNor KCN) to give cyano derivative (v). The halo group of (v) can becoupled to R³-M, where M is a boronic acid, boronic ester or anappropriately substituted metal (e.g., R³-M is R³—B(OH)₂, R³—Sn(Bu)₄, orZn—R³), under standard Suzuki conditions or standard Stille conditions(e.g., in the presence of a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0) and a base (e.g., a bicarbonateor carbonate base) or standard Negishi conditions (e.g., in the presenceof a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0)), to give a derivative offormula (vi). Alternatively, R³-M can be a cyclic amine (where M is Hand attached to the amine nitrogen) and coupled to compound (v) byheating in base or under Buchwald conditions (e.g., in the presence of apalladium(0) catalyst, such as tetrakis(triphenylphosphine)palladium(0)and a base (e.g., an alkoxide base)) to afford ketone (vi). Reduction ofthe ketone (vi) with a suitable reagent, such as sodium tetrahydroborateor the Corey CBS reagent can furnish the alcohol which can be convertedto a derivative bearing a leaving group, (e.g., Lg is chloride viareaction with cyanuric chloride or mesylate via reaction withmethanesulfonic anhydride) and then converted to amine (vii) via theazide. Amine (vii) can be converted to Formula I (viii) using conditionsdescribed in Schemes I, II and III. Alternatively the sequence can beinverted so that ketone (v) can be converted to amine (ix) and (x) andthen the Suzuki, Stille, Negishi or Buchwald coupling is performed togive compounds of Formula I (viii) using conditions described in SchemesI, II and III.

Compounds of Formula I can also be formed as shown in Scheme XVI.Compound (i) can be acylated with a suitable acylating reagent (e.g.,R¹—C(═O)Cl) to form an ester which can be rearranged under Lewis acidconditions (e.g., AlCl₃ or BF₃/HOAc complex) to afford ketone (ii).Halogenation of ketone (ii) using NX¹S (e.g., NX¹S=N-chlorosuccinamide,N-bromosuccinamide or N-iodosuccinamide) can give compound (iii), whereX¹═Cl, Br, or I. The phenol can be converted to an ether (iv) usingstandard conditions (e.g., inorganic base, such as K₂CO₃, and an alkylhalide, such as Et-I). The halo group of (iv) can be coupled to R³-M,where M is a boronic acid, boronic ester or an appropriately substitutedmetal (e.g., R³-M is R³—B(OH)₂, R³—Sn(Bu)₄, or Zn—R³ and R³ is asubstituted or unsubstituted olefin, such as vinyl) under standardSuzuki conditions or standard Stille conditions (e.g., in the presenceof a palladium(0) catalyst, such astetrakis(triphenylphosphine)palladium(0) and a base (e.g., a bicarbonateor carbonate base) to give a derivative of formula (v). The alkene canthen be dihydroxylated using Sharpless conditions to afford the diol(vi). Enhancement of one enantiomer of the secondary alcohol can beachieved using standard Sharpless asymmetric dihydroxylation methods.The secondary alcohol can be converted to the N-Boc protected amine viaa 6-step process (e.g. silyl protection (e.g., TBS-Cl and DIEA) of theprimary alcohol, mesylation of the secondary alcohol, displacement ofthe mesylate with NaN₃, reduction of the azide with Ph₃P, Boc protectionof the resulting primary amine and then deprotection of the silylprotecting group on the primary alcohol with TBAF) to affordamino-alcohol (vii). The amino-alcohol (vii) can be converted into theoxazolidinone by treatment with phosgene to give ketone (viii). Ketone(viii) can be converted to compounds of Formula I (x) using conditionsdescribed in Schemes I, II and III.

Methods

The compounds of the invention can modulate activity of one or more ofvarious kinases including, for example, phosphoinositide 3-kinases(PI3Ks). The term “modulate” is meant to refer to an ability to increaseor decrease the activity of one or more members of the PI3K family.Accordingly, the compounds of the invention can be used in methods ofmodulating a PI3K by contacting the PI3K with any one or more of thecompounds or compositions described herein. In some embodiments,compounds of the present application can act as inhibitors of one ormore PI3Ks. In further embodiments, the compounds of the invention canbe used to modulate activity of a PI3K in an individual in need ofmodulation of the receptor by administering a modulating amount of acompound of the invention, or a pharmaceutically acceptable saltthereof. In some embodiments, modulating is inhibiting.

Given that cancer cell growth and survival is impacted by multiplesignaling pathways, the present application is useful for treatingdisease states characterized by drug resistant kinase mutants. Inaddition, different kinase inhibitors, exhibiting different preferencesin the kinases which they modulate the activities of, may be used incombination. This approach could prove highly efficient in treatingdisease states by targeting multiple signaling pathways, reduce thelikelihood of drug-resistance arising in a cell, and reduce the toxicityof treatments for disease.

Kinases to which the present compounds bind and/or modulate (e.g.,inhibit) include any member of the PI3K family. In some embodiments, thePI3K is PI3Kα, PI3Kβ, PI3Kγ, or PI3Kδ. In some embodiments, the PI3K isPI3Kγ or PI3Kδ. In some embodiments, the PI3K is PI3Kγ. In someembodiments, the PI3K is PI3Kδ. In some embodiments, the PI3K includes amutation. A mutation can be a replacement of one amino acid for another,or a deletion of one or more amino acids. In such embodiments, themutation can be present in the kinase domain of the PI3K.

In some embodiments, more than one compound of the invention is used toinhibit the activity of one kinase (e.g., PI3Kγ or PI3Kδ).

In some embodiments, more than one compound of the invention is used toinhibit more than one kinase, such as at least two kinases (e.g., PI3Kγand PI3Kδ).

In some embodiments, one or more of the compounds is used in combinationwith another kinase inhibitor to inhibit the activity of one kinase(e.g., PI3Kγ or PI3Kδ).

In some embodiments, one or more of the compounds is used in combinationwith another kinase inhibitor to inhibit the activities of more than onekinase (e.g., PI3Kγ or PI3Kδ), such as at least two kinases.

The compounds of the invention can be selective. By “selective” is meantthat the compound binds to or inhibits a kinase with greater affinity orpotency, respectively, compared to at least one other kinase. In someembodiments, the compounds of the invention are selective inhibitors ofPI3Kγ or PI3Kδ over PI3Kα and/or PI3Kβ. In some embodiments, thecompounds of the invention are selective inhibitors of PI3Kδ (e.g., overPI3Kα, PI3Kβ and PI3Kγ). In some embodiments, the compounds of theinvention are selective inhibitors of PI3Kγ (e.g., over PI3Kα, PI3Kβ andPI3Kδ). In some embodiments, selectivity can be at least about 2-fold,5-fold, 10-fold, at least about 20-fold, at least about 50-fold, atleast about 100-fold, at least about 200-fold, at least about 500-foldor at least about 1000-fold. Selectivity can be measured by methodsroutine in the art. In some embodiments, selectivity can be tested atthe K_(m) ATP concentration of each enzyme. In some embodiments, theselectivity of compounds of the invention can be determined by cellularassays associated with particular PI3K kinase activity.

Another aspect of the present invention pertains to methods of treatinga kinase (such as PI3K)-associated disease or disorder in an individual(e.g., patient) by administering to the individual in need of suchtreatment a therapeutically effective amount or dose of one or morecompounds of the present application or a pharmaceutical compositionthereof. A PI3K-associated disease can include any disease, disorder orcondition that is directly or indirectly linked to expression oractivity of the PI3K, including overexpression and/or abnormal activitylevels. In some embodiments, the disease can be linked to Akt (proteinkinase B), mammalian target of rapamycin (mTOR), orphosphoinositide-dependent kinase 1 (PDK1). In some embodiments, themTOR-related disease can be inflammation, atherosclerosis, psoriasis,restenosis, benign prostatic hypertrophy, bone disorders, pancreatitis,angiogenesis, diabetic retinopathy, atherosclerosis, arthritis,immunological disorders, kidney disease, or cancer. A PI3K-associateddisease can also include any disease, disorder or condition that can beprevented, ameliorated, or cured by modulating PI3K activity. In someembodiments, the disease is characterized by the abnormal activity ofPI3K. In some embodiments, the disease is characterized by mutant PI3K.In such embodiments, the mutation can be present in the kinase domain ofthe PI3K.

Examples of PI3K-associated diseases include immune-based diseasesinvolving the system including, for example, rheumatoid arthritis,allergy, asthma, glomerulonephritis, lupus, or inflammation related toany of the above.

Further examples of PI3K-associated diseases include cancers such asbreast, prostate, colon, endometrial, brain, bladder, skin, uterus,ovary, lung, pancreatic, renal, gastric, or hematological cancer.

Further examples of PI3K-associated diseases include lung diseases suchas acute lung injury (ALI) and adult respiratory distress syndrome(ARDS).

Further examples of PI3K-associated diseases include osteoarthritis,restenosis, atherosclerosis, bone disorders, arthritis, diabeticretinopathy, psoriasis, benign prostatic hypertrophy, inflammation,angiogenesis, pancreatitis, kidney disease, inflammatory bowel disease,myasthenia gravis, multiple sclerosis, or Sjögren's syndrome, and thelike.

Further examples of PI3K-associated diseases include idiopathicthrombocytopenic purpura (ITP), autoimmune hemolytic anemia (AIHA),vasculitis, systemic lupus erythematosus, lupus nephritis, pemphigus,membranous nephropathy, chronic lymphocytic leukemia (CLL), Non-Hodgkinlymphoma, hairy cell leukemia, Mantle cell lymphoma, Burkitt lymphoma,small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacyticlymphoma, extranodal marginal zone lymphoma, activated B-cell like (ABC)diffuse large B cell lymphoma, or germinal center B cell (GCB) diffuselarge B cell lymphoma.

In some embodiments, the present application provides a method oftreating pemphigus, membranous nephropathy, Hodgkin's lymphoma,Waldenstrom's macroglobulinemia, prolymphocytic leukemia, acutelymphoblastic leukemia, myelofibrosis, mucosa-associated lymphatictissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma,lymphomatoid granulomatosis, splenic marginal zone lymphoma, primaryeffusion lymphoma, intravascular large B-cell lymphoma, plasma cellleukemia, extramedullary plasmacytoma, smouldering myeloma (akaasymptomatic myeloma), or monoclonal gammopathy of undeterminedsignificance (MGUS).

In some embodiments, the present application provides a method oftreating osteoarthritis, restenosis, atherosclerosis, bone disorders,arthritis, diabetic retinopathy, psoriasis, benign prostatichypertrophy, inflammation, angiogenesis, pancreatitis, kidney disease,inflammatory bowel disease, myasthenia gravis, multiple sclerosis, orSjögren's syndrome.

In some embodiments, the disease is idiopathic thrombocytopenic purpura(ITP), autoimmune hemolytic anemia (AIHA), vasculitis, pemphigus, ormembranous nephropathy.

In some embodiments, the idiopathic thrombocytopenic purpura (ITP) isselected from relapsed ITP and refractory ITP.

In some embodiments, the vasculitis is selected from Behçet's disease,Cogan's syndrome, giant cell arteritis, polymyalgia rheumatica (PMR),Takayasu's arteritis, Buerger's disease (thromboangiitis obliterans),central nervous system vasculitis, Kawasaki disease, polyarteritisnodosa, Churg-Strauss syndrome, mixed cryoglobulinemia vasculitis(essential or hepatitis C virus (HCV)-induced), Henoch-Schönlein purpura(HSP), hypersensitivity vasculitis, microscopic polyangiitis, Wegener'sgranulomatosis, and anti-neutrophil cytoplasm antibody associated (ANCA)systemic vasculitis (AASV).

In some embodiments, the present application provides methods oftreating an immune-based disease, cancer, or lung disease in a patient.

In some embodiments, the immune-based disease is systemic lupuserythematosus or lupus nephritis.

In some embodiments, the cancer is breast cancer, prostate cancer, coloncancer, endometrial cancer, brain cancer, bladder cancer, skin cancer,cancer of the uterus, cancer of the ovary, lung cancer, pancreaticcancer, renal cancer, gastric cancer, or a hematological cancer.

In some embodiments, the hematological cancer is acute myeloblasticleukemia, chronic myeloid leukemia, B cell lymphoma, chronic lymphocyticleukemia (CLL), Non-Hodgkins lymphoma, hairy cell leukemia, Mantle celllymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicularlymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma,activated B-cell like (ABC) diffuse large B cell lymphoma, or germinalcenter B cell (GCB) diffuse large B cell lymphoma.

In some embodiments, the non-Hodgkin lymphoma (NHL) is selected fromrelapsed NHL, refractory NHL, and recurrent follicular NHL.

In some embodiments, the lung disease is acute lung injury (ALI) oradult respiratory distress syndrome (ARDS).

As used herein, the term “contacting” refers to the bringing together ofindicated moieties in an in vitro system or an in vivo system. Forexample, “contacting” a PI3K with a compound of the invention includesthe administration of a compound of the present application to anindividual or patient, such as a human, having a PI3K, as well as, forexample, introducing a compound of the invention into a samplecontaining a cellular or purified preparation containing the PI3K.

As used herein, the term “individual” or “patient,” usedinterchangeably, refers to any animal, including mammals, preferablymice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep,horses, or primates, and most preferably humans.

As used herein, the phrase “therapeutically effective amount” refers tothe amount of active compound or pharmaceutical agent that elicits thebiological or medicinal response that is being sought in a tissue,system, animal, individual or human by a researcher, veterinarian,medical doctor or other clinician. In some embodiments, the dosage ofthe compound, or a pharmaceutically acceptable salt thereof,administered to a patient or individual is about 1 mg to about 2 g,about 1 mg to about 1000 mg, about 1 mg to about 500 mg, about 1 mg toabout 100 mg, about 1 mg to 50 mg, or about 50 mg to about 500 mg.

As used herein, the term “treating” or “treatment” refers to one or moreof (1) preventing the disease; for example, preventing a disease,condition or disorder in an individual who may be predisposed to thedisease, condition or disorder but does not yet experience or displaythe pathology or symptomatology of the disease; (2) inhibiting thedisease; for example, inhibiting a disease, condition or disorder in anindividual who is experiencing or displaying the pathology orsymptomatology of the disease, condition or disorder (i.e., arrestingfurther development of the pathology and/or symptomatology); and (3)ameliorating the disease; for example, ameliorating a disease, conditionor disorder in an individual who is experiencing or displaying thepathology or symptomatology of the disease, condition or disorder (i.e.,reversing the pathology and/or symptomatology) such as decreasing theseverity of disease.

Combination Therapies

One or more additional pharmaceutical agents such as, for example,chemotherapeutics, anti-inflammatory agents, steroids,immunosuppressants, as well as Bcr-Abl, Flt-3, EGFR, HER2, JAK (e.g.,JAK1 or JAK2), c-MET, VEGFR, PDGFR, cKit, IGF-1R, RAF, FAK, Akt mTOR,PIM, and AKT (e.g., AKT1, AKT2, or AKT3) kinase inhibitors such as, forexample, those described in WO 2006/056399, or other agents such as,therapeutic antibodies can be used in combination with the compounds ofthe present application for treatment of PI3K-associated diseases,disorders or conditions. The one or more additional pharmaceuticalagents can be administered to a patient simultaneously or sequentially.

In some embodiments, the additional pharmaceutical agent is a JAK1and/or JAK2 inhibitor. In some embodiments, the present applicationprovides a method of treating a disease described herein (e.g., a B cellmalignancy, such as diffuse B-cell lymphoma) in a patient comprisingadministering to the patient a compound described herein, or apharmaceutically acceptable salt thereof, and a JAK1 and/or JAK2inhibitor. The B cell malignancies can include those described hereinand in U.S. Ser. No. 61/976,815, filed April 8, 2014. In someembodiments, the inhibitor of JAK1 and/or JAK2 is3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile.In some embodiments, the inhibitor of JAK1 and/or JAK2 is(3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile(ruxolitinib; also known as INCB018424). Ruxolitinib has an IC₅₀ of lessthan 10 nM at 1 mM ATP (assay J) at JAK1 and JAK2. 3-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrileand ruxolitinib can be made by the procedure described in U.S. Pat. No.7,598,257 (Example 67), filed Dec. 12, 2006, which is incorporatedherein by reference in its entirety. In some embodiments, the inhibitorof JAK1 and/or JAK2 is(3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrilephosphoric acid salt.

In some embodiments, the inhibitor of JAK1 and/or JAK2 is a compound ofTable 1, or a pharmaceutically acceptable salt thereof. The compounds inTable 1 are selective JAK1 inhibitors (selective over JAK2, JAK3, andTYK2). The IC₅₀s obtained by the method of Assay J at 1 mM ATP are shownin Table 1.

TABLE 1 JAK1 IC₅₀ JAK2/ # Prep. Name Structure (nM) JAK1 1 Example J1herein ((2R,5S)-5-{2-[(1R)- 1-hydroxyethyl]-1H- imidazo[4,5-d]thieno[3,2- b]pyridin-1- yl}tetrahydro-2H- pyran-2-yl)acetonitrile

++ >10 2 Example J2 herein 4-[3-(cyanomethyl)-3- (3′,5′-dimethyl-1H,1′H-4,4′-bipyrazol- 1-yl)azetidin-1-yl]- 2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1- methylethyl]benzamide

+++ >10 3 US 2010/ 0298334 (Example 2)^(a) 3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]- 3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H-pyrazol-1- yl]propanenitrile

+ >10 4 US 2010/ 0298334 (Example 13c) 3-(1-[1,3]oxazolo[5,4-b]pyridin-2- ylpyrrolidin-3-yl)-3- [4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H- pyrazol-1- yl]propanenitrile

+ >10 5 US 2011/ 0059951 (Example 12) 4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1- yl]propyl}piperazin-1-yl)carbonyl]-3- fluorobenzonitrile

+ >10 6 US 2011/ 0059951 (Example 13) 4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrrol-1- yl]propyl}piperazin-1-yl)carbonyl]-3- fluorobenzonitrile

+ >10 7 US 2011/ 0224190 (Example 1) {1-{1-[3-Fluoro-2-(trifluoromethyl)iso- nicotinoyl]piperidin-4- yl}-3-[4-(7H- pyrrolo[2,3-d]pyrimidin-4-yl)-1H- pyrazol-1-yl]azetidin- 3-yl}acetonitrile

+ >10 8 US 2011/ 0224190 (Example 154) 4-{3-(Cyanomethyl)-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1-yl]azetidin-1-yl}-N-[4-fluoro-2- (trifluoromethyl) phenyl]piperidine-1- carboxamide

+ >10 9 US 2011/ 0224190 (Example 85) [3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H- pyrazol-1-yl]-1-(1- {[2- (trifluoromethyl)pyrimidin-4- yl]carbonyl}piperidin- 4-yl)azetidin-3- yl]acetonitrile

+ >10 10 US 2012/ 0149681 (Example 7b) [trans-1-[4-(7H- pyrrolo[2,3-d]pyrimidin-4-yl)-1H- pyrazol-1-yl]-3-(4- {[2- (trifluoromethyl)pyrimidin-4- yl]carbonyl}piperazin- 1- yl)cyclobutyl] acetonitrile

+ >10 11 US 2012/ 0149681 (Example 157) {trans-3-(4-{[4-[(3-hydroxyazetidin-1- yl)methyl]-6- (trifluoromethyl) pyridin-2-yl]oxy}piperidin-1-yl)-1-[4- (7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1-yl]cyclobutyl} acetonitrile

+ >10 12 US 2012/ 0149681 (Example 161) {trans-3-(4-{[4- {[(2S)-2-(hydroxymethyl) pyrrolidin-1-yl]methyl}- 6-(trifluoromethyl)pyridin-2-yl]oxy} piperidin-1-yl)-1-[4- (7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H- pyrazol-1- yl]cyclobutyl} acetonitrile

+ >10 13 US 2012/ 0149681 (Example 162) {trans-3-(4-{[4- {[(2R)-2-(hydroxymethyl) pyrrolidin-1-yl]methyl}- 6-(trifluoromethyl)pyridin-2-yl]oxy} piperidin-1-yl)-1-[4- (7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H- pyrazol-1- yl]cyclobutyl} acetonitrile

+ >10 14 US 2012/ 0149682 (Example 20)^(b) 4-(4-{3- [(dimethylamino)methyl]-5- fluorophenoxy} piperidin-1-yl)-3-[4- (7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H- pyrazol-1- yl]butanenitrile

+ >10 15 US 2013/ 0018034 (Example 18) 5-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1-yl]azetidin-1-yl}-N- isopropylpyrazine-2- carboxamide

+ >10 16 US 2013/ 0018034 (Example 28) 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N- [(1S)-2,2,2-trifluoro-1- methylethyl]benzamide

+ >10 17 US 2013/ 0018034 (Example 34) 5-{3-(cyanomethyl)-3-[4-(1H-pyrrolo[2,3- b]pyridin-4-yl)-1H- pyrazol-1-yl]azetidin- 1-yl}-N-isopropylpyrazine-2- carboxamide

+ >10 18 US 2013/ 0045963 (Example 45) {1-(cis-4-{[6-(2-hydroxyethyl)-2- (trifluoromethyl) pyrimidin-4- yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1-yl]azetidin-3-yl}acetonitrile

+ >10 19 US 2013/ 0045963 (Example 65) {1-(cis-4-{[4-[(ethylamino)methyl]- 6- (trifluoromethyl) pyridin-2-yl]oxy}cyclohexyl)-3- [4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin- 3-yl}acetonitrile

+ >10 20 US 2013/ 0045963 (Example 69) {1-(cis-4-{[4-(1- hydroxy-1-methylethyl)-6- (trifluoromethyl) pyridin-2- yl]oxy}cyclohexyl)-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1-yl]azetidin-3-yl}acetonitrile

+ >10 21 US 2013/ 0045963 (Example 95) {1-(cis-4-{[4-{[(3R)-3-hydroxypyrrolidin- 1-yl]methyl}-6- (trifluoromethyl) pyridin-2-yl]oxy}cyclohexyl)-3- [4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin- 3-yl}acetonitrile

+ >10 22 US 2013/ 0045963 (Example 95) {1-(cis-4-{[4-{[(3S)-3-hydroxypyrrolidin- 1-yl]methyl}-6- (trifluoromethyl) pyridin-2-yl]oxy}cyclohexyl)-3- [4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin- 3-yl}acetonitrile

+ >10 23 US 2014/ 0005166 (Example 1) {trans-3-(4-{[4-({[(1S)-2-hydroxy-1- methylethyl]amino} methyl)-6- (trifluoromethyl)pyridin-2-yl]oxy} piperidin-1-yl)-1-[4- (7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H- pyrazol-1- yl]cyclobutyl} acetonitrile

+ >10 24 US 2014/ 0005166 (Example 14) {trans-3-(4-{[4- ({[(2R)-2-hydroxypropyl]amino} methyl)-6- (trifluoromethyl) pyridin-2-yl]oxy}piperidin-1-yl)-1-[4- (7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1-yl]cyclobutyl} acetonitrile

+ >10 25 US 2014/ 0005166 (Example 15) {trans-3-(4-{[4- ({[(2S)-2-hydroxypropyl]amino} methyl)-6- (trifluoromethyl) pyridin-2-yl]oxy}piperidin-1-yl)-1-[4- (7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1-yl]cyclobutyl} acetonitrile

+ >10 26 US 2014/ 0005166 (Example 20) {trans-3-(4-{[4-(2-hydroxyethyl)-6- (trifluoromethyl) pyridin-2-yl]oxy}piperidin-1-yl)-1-[4- (7H-pyrrolo[2,3- d]pyrimidin-4-yl)-1H- pyrazol-1-yl]cyclobutyl} acetonitrile

+ >10 +means <10 nM (see Example A for assay conditions) ++means <100 nM(see Example A for assay conditions) +++means <300 nM (see Example A forassay conditions) ^(a)Data for enantiomer 1 ^(b)Data for enantiomer 2

In some embodiments, the inhibitor of JAK1 and/or JAK2 is{1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile,or a pharmaceutically acceptable salt thereof.

In some embodiments, the inhibitor of JAK1 and/or JAK2 is{1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrileadipic acid salt.

In some embodiments, the inhibitor of JAK1 and/or JAK2 is4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide,or a pharmaceutically acceptable salt thereof.

In some embodiments, the inhibitor of JAK1 and/or JAK2 is selected from(R)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,(R)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,(R)-4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile,(R)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile,or(R)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile,(S)-3-[1-(6-chloropyridin-2-yl)pyrrolidin-3-yl]-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,(S)-3-(1-[1,3]oxazolo[5,4-b]pyridin-2-ylpyrrolidin-3-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,(S)-4-[(4-{3-cyano-2-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile,(S)-4-[(4-{3-cyano-2-[3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrol-1-yl]propyl}piperazin-1-yl)carbonyl]-3-fluorobenzonitrile,(S)-4-(4-{3-[(dimethylamino)methyl]-5-fluorophenoxy}piperidin-1-yl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile;and pharmaceutically acceptable salts of any of the aforementioned.

In some embodiments, the compounds of Table 1 are prepared by thesynthetic procedures described in US Patent Publ. No. 2010/0298334,filed May 21, 2010, US Patent Publ. No. 2011/0059951, filed Aug. 31,2010, US Patent Publ. No. 2011/0224190, filed Mar. 9, 2011, US PatentPubl. No. 2012/0149681, filed Nov. 18, 2011, US Patent Publ. No.2012/0149682, filed Nov. 18, 2011, US Patent Publ. 2013/0018034, filedJun. 19, 2012, US Patent Publ. No. 2013/0045963, filed Aug. 17, 2012,and US Patent Publ. No. 2014/0005166, filed May 17, 2013, each of whichis incorporated herein by reference in its entirety.

In some embodiments, the inhibitor of JAK1 and/or JAK2 is selected fromthe compounds of US Patent Publ. No. 2010/0298334, filed May 21, 2010,US Patent Publ. No. 2011/0059951, filed Aug. 31, 2010, US Patent Publ.No. 2011/0224190, filed Mar. 9, 2011, US Patent Publ. No. 2012/0149681,filed Nov. 18, 2011, US Patent Publ. No. 2012/0149682, filed Nov. 18,2011, US Patent Publ. 2013/0018034, filed Jun. 19, 2012, US Patent Publ.No. 2013/0045963, filed Aug. 17, 2012, and US Patent Publ. No.2014/0005166, filed May 17, 2013, each of which is incorporated hereinby reference in its entirety.

Example antibodies for use in combination therapy include but are notlimited to Trastuzumab (e.g. anti-HER2), Ranibizumab (e.g. anti-VEGF-A),Bevacizumab (trade name Avastin, e.g. anti-VEGF, Panitumumab (e.g.anti-EGFR), Cetuximab (e.g. anti-EGFR), Rituxan (anti-CD20) andantibodies directed to c-MET.

One or more of the following agents may be used in combination with thecompounds of the present application and are presented as a non-limitinglist: a cytostatic agent, cisplatin, doxorubicin, taxotere, taxol,etoposide, irinotecan, camptostar, topotecan, paclitaxel, docetaxel,epothilones, tamoxifen, 5-fluorouracil, methoxtrexate, temozolomide,cyclophosphamide, SCH 66336, R115777, L778,123, BMS 214662, Iressa,Tarceva, antibodies to EGFR, Gleevec™, intron, ara-C, adriamycin,cytoxan, gemcitabine, Uracil mustard, Chlormethine, Ifosfamide,Melphalan, Chlorambucil, Pipobroman, Triethylenemelamine,Triethylenethiophosphoramine, Busulfan, Carmustine, Lomustine,Streptozocin, Dacarbazine, Floxuridine, Cytarabine, 6-Mercaptopurine,6-Thioguanine, Fludarabine phosphate, oxaliplatin, leucovirin,ELOXATIN™, Pentostatine, Vinblastine, Vincristine, Vindesine, Bleomycin,Dactinomycin, Daunorubicin, Doxorubicin, Epirubicin, Idarubicin,Mithramycin, Deoxycoformycin, Mitomycin-C, L-Asparaginase, Teniposide17.alpha.-Ethinylestradiol, Diethylstilbestrol, Testosterone,Prednisone, Fluoxymesterone, Dromostanolone propionate, Testolactone,Megestrolacetate, Methylprednisolone, Methyltestosterone, Prednisolone,Triamcinolone, Chlorotrianisene, Hydroxyprogesterone, Aminoglutethimide,Estramustine, Medroxyprogesteroneacetate, Leuprolide, Flutamide,Toremifene, goserelin, Cisplatin, Carboplatin, Hydroxyurea, Amsacrine,Procarbazine, Mitotane, Mitoxantrone, Levamisole, Navelbene,Anastrazole, Letrazole, Capecitabine, Reloxafine, Droloxafine,Hexamethylmelamine, Avastin, herceptin, Bexxar, Velcade, Zevalin,Trisenox, Xeloda, Vinorelbine, Porfimer, Erbitux, Liposomal, Thiotepa,Altretamine, Melphalan, Trastuzumab, Lerozole, Fulvestrant, Exemestane,Fulvestrant, Ifosfomide, Rituximab, C225, Campath, Clofarabine,cladribine, aphidicolon, rituxan, sunitinib, dasatinib, tezacitabine,Sml1, fludarabine, pentostatin, triapine, didox, trimidox, amidox, 3-AP,MDL-101,731, bendamustine (Treanda), ofatumumab, and GS-1101 (also knownas CAL-101).

Example chemotherapeutics include proteosome inhibitors (e.g.,bortezomib), thalidomide, revlimid, and DNA-damaging agents such asmelphalan, doxorubicin, cyclophosphamide, vincristine, etoposide,carmustine, and the like.

Example steroids include coriticosteroids such as dexamethasone orprednisone.

Example Bcr-Abl inhibitors include the compounds, and pharmaceuticallyacceptable salts thereof, of the genera and species disclosed in U.S.Pat. No. 5,521,184, WO 04/005281, and U.S. Ser. No. 60/578,491.

Example suitable Flt-3 inhibitors include compounds, and theirpharmaceutically acceptable salts, as disclosed in WO 03/037347, WO03/099771, and WO 04/046120.

Example suitable RAF inhibitors include compounds, and theirpharmaceutically acceptable salts, as disclosed in WO 00/09495 and WO05/028444.

Example suitable FAK inhibitors include compounds, and theirpharmaceutically acceptable salts, as disclosed in WO 04/080980, WO04/056786, WO 03/024967, WO 01/064655, WO 00/053595, and WO 01/014402.

Example suitable mTOR inhibitors include compounds, and theirpharmaceutically acceptable salts, as disclosed in WO 2011/025889.

In some embodiments, the compounds of the invention can be used incombination with one or more other kinase inhibitors including imatinib,particularly for treating patients resistant to imatinib or other kinaseinhibitors.

In some embodiments, the compounds of the invention can be used incombination with a chemotherapeutic in the treatment of cancer, such asmultiple myeloma, and may improve the treatment response as compared tothe response to the chemotherapeutic agent alone, without exacerbationof its toxic effects. Examples of additional pharmaceutical agents usedin the treatment of multiple myeloma, for example, can include, withoutlimitation, melphalan, melphalan plus prednisone [MP], doxorubicin,dexamethasone, and Velcade (bortezomib). Further additional agents usedin the treatment of multiple myeloma include Bcr-Abl, Flt-3, RAF and FAKkinase inhibitors. Additive or synergistic effects are desirableoutcomes of combining a PI3K inhibitor of the present application withan additional agent. Furthermore, resistance of multiple myeloma cellsto agents such as dexamethasone may be reversible upon treatment withthe PI3K inhibitor of the present application. The agents can becombined with the present compound in a single or continuous dosageform, or the agents can be administered simultaneously or sequentiallyas separate dosage forms.

In some embodiments, a corticosteroid such as dexamethasone isadministered to a patient in combination with the compounds of theinvention where the dexamethasone is administered intermittently asopposed to continuously.

In some further embodiments, combinations of the compounds of theinvention with other therapeutic agents can be administered to a patientprior to, during, and/or after a bone marrow transplant or stem celltransplant.

Pharmaceutical Formulations and Dosage Forms

When employed as pharmaceuticals, the compounds of the invention can beadministered in the form of pharmaceutical compositions. Thesecompositions can be prepared in a manner well known in thepharmaceutical art, and can be administered by a variety of routes,depending upon whether local or systemic treatment is desired and uponthe area to be treated. Administration may be topical (includingtransdermal, epidermal, ophthalmic and to mucous membranes includingintranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalationor insufflation of powders or aerosols, including by nebulizer;intratracheal or intranasal), oral or parenteral. Parenteraladministration includes intravenous, intraarterial, subcutaneous,intraperitoneal intramuscular or injection or infusion; or intracranial,e.g., intrathecal or intraventricular, administration. Parenteraladministration can be in the form of a single bolus dose, or may be, forexample, by a continuous perfusion pump. Pharmaceutical compositions andformulations for topical administration may include transdermal patches,ointments, lotions, creams, gels, drops, suppositories, sprays, liquidsand powders. Conventional pharmaceutical carriers, aqueous, powder oroily bases, thickeners and the like may be necessary or desirable.

This invention also includes pharmaceutical compositions which contain,as the active ingredient, the compound of the invention or apharmaceutically acceptable salt thereof, in combination with one ormore pharmaceutically acceptable carriers (excipients). In someembodiments, the composition is suitable for topical administration. Inmaking the compositions of the invention, the active ingredient istypically mixed with an excipient, diluted by an excipient or enclosedwithin such a carrier in the form of, for example, a capsule, sachet,paper, or other container. When the excipient serves as a diluent, itcan be a solid, semi-solid, or liquid material, which acts as a vehicle,carrier or medium for the active ingredient. Thus, the compositions canbe in the form of tablets, pills, powders, lozenges, sachets, cachets,elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solidor in a liquid medium), ointments containing, for example, up to 10% byweight of the active compound, soft and hard gelatin capsules,suppositories, sterile injectable solutions, and sterile packagedpowders. In preparing a formulation, the active compound can be milledto provide the appropriate particle size prior to combining with theother ingredients. If the active compound is substantially insoluble, itcan be milled to a particle size of less than 200 mesh. If the activecompound is substantially water soluble, the particle size can beadjusted by milling to provide a substantially uniform distribution inthe formulation, e.g. about 40 mesh.

The compounds of the invention may be milled using known millingprocedures such as wet milling to obtain a particle size appropriate fortablet formation and for other formulation types. Finely divided(nanoparticulate) preparations of the compounds of the invention can beprepared by processes known in the art, e.g., see International App. No.WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. Theformulations can additionally include: lubricating agents such as talc,magnesium stearate, and mineral oil; wetting agents; emulsifying andsuspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; sweetening agents; and flavoring agents. Thecompositions of the invention can be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.

The compositions can be formulated in a unit dosage form, each dosagecontaining from about 5 to about 1000 mg (1 g), more usually about 100to about 500 mg, of the active ingredient. The term “unit dosage forms”refers to physically discrete units suitable as unitary dosages forhuman subjects and other mammals, each unit containing a predeterminedquantity of active material calculated to produce the desiredtherapeutic effect, in association with a suitable pharmaceuticalexcipient.

In some embodiments, the compositions of the invention contain fromabout 5 to about 50 mg of the active ingredient. One having ordinaryskill in the art will appreciate that this embodies compositionscontaining about 5 to about 10, about 10 to about 15, about 15 to about20, about 20 to about 25, about 25 to about 30, about 30 to about 35,about 35 to about 40, about 40 to about 45, or about 45 to about 50 mgof the active ingredient.

In some embodiments, the compositions of the invention contain fromabout 50 to about 500 mg of the active ingredient. One having ordinaryskill in the art will appreciate that this embodies compositionscontaining about 50 to about 100, about 100 to about 150, about 150 toabout 200, about 200 to about 250, about 250 to about 300, about 350 toabout 400, or about 450 to about 500 mg of the active ingredient.

In some embodiments, the compositions of the invention contain fromabout 500 to about 1000 mg of the active ingredient. One having ordinaryskill in the art will appreciate that this embodies compositionscontaining about 500 to about 550, about 550 to about 600, about 600 toabout 650, about 650 to about 700, about 700 to about 750, about 750 toabout 800, about 800 to about 850, about 850 to about 900, about 900 toabout 950, or about 950 to about 1000 mg of the active ingredient.

Similar dosages may be used of the compounds described herein in themethods and uses of the invention.

The active compound can be effective over a wide dosage range and isgenerally administered in a pharmaceutically effective amount. It willbe understood, however, that the amount of the compound actuallyadministered will usually be determined by a physician, according to therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered, theage, weight, and response of the individual patient, the severity of thepatient's symptoms, and the like.

For preparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical excipient to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present application. When referring to thesepreformulation compositions as homogeneous, the active ingredient istypically dispersed evenly throughout the composition so that thecomposition can be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid preformulation isthen subdivided into unit dosage forms of the type described abovecontaining from, for example, about 0.1 to about 1000 mg of the activeingredient of the present application.

The tablets or pills of the present application can be coated orotherwise compounded to provide a dosage form affording the advantage ofprolonged action. For example, the tablet or pill can comprise an innerdosage and an outer dosage component, the latter being in the form of anenvelope over the former. The two components can be separated by anenteric layer which serves to resist disintegration in the stomach andpermit the inner component to pass intact into the duodenum or to bedelayed in release. A variety of materials can be used for such entericlayers or coatings, such materials including a number of polymeric acidsand mixtures of polymeric acids with such materials as shellac, cetylalcohol, and cellulose acetate.

The liquid forms in which the compounds and compositions of the presentapplication can be incorporated for administration orally or byinjection include aqueous solutions, suitably flavored syrups, aqueousor oil suspensions, and flavored emulsions with edible oils such ascottonseed oil, sesame oil, coconut oil, or peanut oil, as well aselixirs and similar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. In some embodiments, the compositions are administered by theoral or nasal respiratory route for local or systemic effect.Compositions can be nebulized by use of inert gases. Nebulized solutionsmay be breathed directly from the nebulizing device or the nebulizingdevice can be attached to a face mask, tent, or intermittent positivepressure breathing machine. Solution, suspension, or powder compositionscan be administered orally or nasally from devices which deliver theformulation in an appropriate manner.

Topical formulations can contain one or more conventional carriers. Insome embodiments, ointments can contain water and one or morehydrophobic carriers selected from, for example, liquid paraffin,polyoxyethylene alkyl ether, propylene glycol, white Vaseline, and thelike. Carrier compositions of creams can be based on water incombination with glycerol and one or more other components, e.g.glycerinemonostearate, PEG-glycerinemonostearate and cetylstearylalcohol. Gels can be formulated using isopropyl alcohol and water,suitably in combination with other components such as, for example,glycerol, hydroxyethyl cellulose, and the like. In some embodiments,topical formulations contain at least about 0.1, at least about 0.25, atleast about 0.5, at least about 1, at least about 2, or at least about 5wt % of the compound of the invention. The topical formulations can besuitably packaged in tubes of, for example, 100 g which are optionallyassociated with instructions for the treatment of the select indication,e.g., psoriasis or other skin condition.

The amount of compound or composition administered to a patient willvary depending upon what is being administered, the purpose of theadministration, such as prophylaxis or therapy, the state of thepatient, the manner of administration, and the like. In therapeuticapplications, compositions can be administered to a patient alreadysuffering from a disease in an amount sufficient to cure or at leastpartially arrest the symptoms of the disease and its complications.Effective doses will depend on the disease condition being treated aswell as by the judgment of the attending clinician depending uponfactors such as the severity of the disease, the age, weight and generalcondition of the patient, and the like.

The compositions administered to a patient can be in the form ofpharmaceutical compositions described above. These compositions can besterilized by conventional sterilization techniques, or may be sterilefiltered. Aqueous solutions can be packaged for use as is, orlyophilized, the lyophilized preparation being combined with a sterileaqueous carrier prior to administration. The pH of the compoundpreparations typically will be between 3 and 11, more preferably from 5to 9 and most preferably from 7 to 8. It will be understood that use ofcertain of the foregoing excipients, carriers, or stabilizers willresult in the formation of pharmaceutical salts.

The therapeutic dosage of a compound of the present application can varyaccording to, for example, the particular use for which the treatment ismade, the manner of administration of the compound, the health andcondition of the patient, and the judgment of the prescribing physician.The proportion or concentration of a compound of the invention in apharmaceutical composition can vary depending upon a number of factorsincluding dosage, chemical characteristics (e.g., hydrophobicity), andthe route of administration. For example, the compounds of the inventioncan be provided in an aqueous physiological buffer solution containingabout 0.1 to about 10% w/v of the compound for parenteraladministration. Some typical dose ranges are from about 1 μg/kg to about1 g/kg of body weight per day. In some embodiments, the dose range isfrom about 0.01 mg/kg to about 100 mg/kg of body weight per day. Thedosage is likely to depend on such variables as the type and extent ofprogression of the disease or disorder, the overall health status of theparticular patient, the relative biological efficacy of the compoundselected, formulation of the excipient, and its route of administration.Effective doses can be extrapolated from dose-response curves derivedfrom in vitro or animal model test systems.

The compositions of the invention can further include one or moreadditional pharmaceutical agents such as a chemotherapeutic, steroid,anti-inflammatory compound, or immunosuppressant, examples of which arelisted herein.

Labeled Compounds and Assay Methods

Another aspect of the present application relates to labeled compoundsof the invention (radio-labeled, fluorescent-labeled, etc.) that wouldbe useful not only in imaging techniques but also in assays, both invitro and in vivo, for localizing and quantitating PI3K in tissuesamples, including human, and for identifying PI3K ligands by inhibitionbinding of a labeled compound. Accordingly, the present applicationincludes PI3K assays that contain such labeled compounds.

The present application further includes isotopically-labeled compoundsof the invention. An “isotopically” or “radio-labeled” compound is acompound of the invention where one or more atoms are replaced orsubstituted by an atom having an atomic mass or mass number differentfrom the atomic mass or mass number typically found in nature (i.e.,naturally occurring). Suitable radionuclides that may be incorporated incompounds of the present application include but are not limited to ³H(also written as T for tritium), 11C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O,¹⁸F, ³⁵S, ³⁶Cl, ⁸²Br, ⁷⁵Br, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I, ¹²⁵I and ¹³¹I. Theradionuclide that is incorporated in the instant radio-labeled compoundswill depend on the specific application of that radio-labeled compound.For example, for in vitro PI3K labeling and competition assays,compounds that incorporate ³H, ¹⁴C, ⁸²Br, ¹²⁵I, ¹³¹I, ³⁵S or willgenerally be most useful. For radio-imaging applications ¹¹C, ¹⁸F, ¹²⁵I,¹²³I, ¹²⁴I, ¹³¹I, ⁷⁵Br, ⁷⁶Br or ⁷⁷Br will generally be most useful.

It is understood that a “radio-labeled” or “labeled compound” is acompound that has incorporated at least one radionuclide. In someembodiments the radionuclide is selected from the group consisting of³H, ¹⁴C, ¹²⁵I, ³⁵S and ⁸²Br. In some embodiments, one or more H atomsfor any compound described herein is each replaced by a deuterium atom.

The present application can further include synthetic methods forincorporating radio-isotopes into compounds of the invention. Syntheticmethods for incorporating radio-isotopes into organic compounds are wellknown in the art, and an ordinary skill in the art will readilyrecognize the methods applicable for the compounds of invention.

A labeled compound of the invention can be used in a screening assay toidentify/evaluate compounds. For example, a newly synthesized oridentified compound (i.e., test compound) which is labeled can beevaluated for its ability to bind a PI3K by monitoring its concentrationvariation when contacting with the PI3K, through tracking of thelabeling. For example, a test compound (labeled) can be evaluated forits ability to reduce binding of another compound which is known to bindto a PI3K (i.e., standard compound). Accordingly, the ability of a testcompound to compete with the standard compound for binding to the PI3Kdirectly correlates to its binding affinity. Conversely, in some otherscreening assays, the standard compound is labeled and test compoundsare unlabeled. Accordingly, the concentration of the labeled standardcompound is monitored in order to evaluate the competition between thestandard compound and the test compound, and the relative bindingaffinity of the test compound is thus ascertained.

Kits

The present application also includes pharmaceutical kits useful, forexample, in the treatment or prevention of PI3K-associated diseases ordisorders, such as cancer, which include one or more containerscontaining a pharmaceutical composition comprising a therapeuticallyeffective amount of a compound of the invention. Such kits can furtherinclude, if desired, one or more of various conventional pharmaceuticalkit components, such as, for example, containers with one or morepharmaceutically acceptable carriers, additional containers, etc., aswill be readily apparent to those skilled in the art. Instructions,either as inserts or as labels, indicating quantities of the componentsto be administered, guidelines for administration, and/or guidelines formixing the components, can also be included in the kit.

The invention will be described in greater detail by way of specificexamples. The following examples are offered for illustrative purposes,and are not intended to limit the invention in any manner. Those ofskill in the art will readily recognize a variety of non-criticalparameters which can be changed or modified to yield essentially thesame results. The compounds of the Examples have been found to be PI3Kinhibitors according to at least one assay described herein.

EXAMPLE S

Experimental procedures for compounds of the invention are providedbelow. The example compounds below containing one or more chiral centerswere obtained in racemate form or as isomeric mixtures, unless otherwisespecified. Salt stoichiometry which is indicated any of the productsbelow is meant only to indicate a probable stoichiometry, and should notbe construed to exclude the possible formation of salts in otherstoichiometries. The abbreviations “h” and “min” refer to hour(s) andminute(s), respectively.

Preparatory LC-MS purifications of some of the compounds prepared wereperformed on Waters mass directed fractionation systems. The basicequipment setup, protocols, and control software for the operation ofthese systems have been described in detail in the literature. See e.g.“Two-Pump At Column Dilution Configuration for Preparative LC-MS”, K.Blom, J. Combi. Chem., 4, 295 (2002); “Optimizing Preparative LC-MSConfigurations and Methods for Parallel Synthesis Purification”, K.Blom, R. Sparks, J. Doughty, G. Everlof, T. Hague, A. Combs, J. Combi.Chem., 5, 670 (2003); and “Preparative LC-MS Purification: ImprovedCompound Specific Method Optimization”, K. Blom, B. Glass, R. Sparks, A.Combs, J. Combi. Chem., 6, 874-883 (2004). The compounds separated weretypically subjected to analytical liquid chromatography massspectrometry (LCMS) for purity analysis under the following conditions:Instrument; Agilent 1100 series, LC/MSD, Column: Waters Sunfire™ C₁₈ 5μm, 2.1×50 mm, Buffers: mobile phase A: 0.025% TFA in water and mobilephase B: acetonitrile; gradient 2% to 80% of B in 3 minutes with flowrate 2.0 mL/minute.

Some of the compounds prepared were also separated on a preparativescale by reverse-phase high performance liquid chromatography (RP-HPLC)with MS detector or flash chromatography (silica gel) as indicated inthe Examples. Typical preparative reverse-phase high performance liquidchromatography (RP-HPLC) column conditions are as follows:

pH=2 purifications: Waters Sunfire™ C₁₈ 5 μm, 19×100 mm column, elutingwith mobile phase A: 0.1% TFA (trifluoroacetic acid) in water and mobilephase B: acetonitrile; the flow rate was 30 mL/minute, the separatinggradient was optimized for each compound using the Compound SpecificMethod Optimization protocol as described in the literature [see“Preparative LCMS Purification: Improved Compound Specific MethodOptimization”, K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem.,6, 874-883 (2004)]. Typically, the flow rate used with the 30×100 mmcolumn was 60 mL/minute.

pH=10 purifications: Waters)(Bridge C₁₈ 5 μm, 19×100 mm column, elutingwith mobile phase A: 0.15% NH₄OH in water and mobile phase B:acetonitrile; the flow rate was 30 mL/minute, the separating gradientwas optimized for each compound using the Compound Specific MethodOptimization protocol as described in the literature [See “PreparativeLCMS Purification: Improved Compound Specific Method Optimization”, K.Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874-883 (2004)].Typically, the flow rate used with 30×100 mm column was 60 mL/minute.

Example 14-{3-Chloro-6-ethoxy-2-fluoro-5-[1-([1,3]thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl]phenyl}pyrrolidin-2-onetrifluoroacetate

Step 1.4-(3-(1-Azidoethyl)-5-chloro-2-ethoxy-6-fluorophenyl)pyrrolidin-2-one

A mixture of4-[3-chloro-6-ethoxy-2-fluoro-5-(1-hydroxyethyl)phenyl]pyrrolidin-2-one(0.437 g, 1.45 mmol) (from US 2013/0059835, Examples 345-348, Step 6)and N,N-dimethylformamide (0.0112 mL, 0.145 mmol) in dichloromethane (12mL) was treated with thionyl chloride (0.222 mL, 3.04 mmol) dropwise.The reaction mixture was stirred for 30 min, added dropwise toice-cooled saturated sodium bicarbonate solution, and extracted withdichloromethane. The organic layer was washed with brine, dried overanhydrous sodium sulfate, filtered, and concentrated to yield a chlorideintermediate (0.463 g, 100%) that was used without further purification.The chloride intermediate was dissolved in N,N-dimethylformamide (4.35mL), treated with sodium azide (0.282 g, 4.34 mmol), and heated at 60°C. for 1 h. The reaction mixture was poured over saturated sodiumbicarbonate solution (30 mL) and water (30 mL) and extracted with ethylacetate (2×50 mL). The combined organic extracts were washed with brine,dried over anhydrous sodium sulfate, filtered, and concentrated to abrown solid. Purification by flash column chromatography (100% DCM to10% MeOH/DCM) afforded the desired product (0.47 g, 99%) as a whitesolid. LCMS calculated for C₁₄H₁₇ClFN₄O₂ (M+H)⁺: m/z=327.1; found:326.9.

Step 2.4-[3-(1-Aminoethyl)-5-chloro-2-ethoxy-6-fluorophenyl]pyrrolidin-2-one

A solution of4-[3-(1-azidoethyl)-5-chloro-2-ethoxy-6-fluorophenyl]pyrrolidin-2-one(0.461 g, 1.41 mmol) in tetrahydrofuran (7.6 mL) and water (1.5 mL) wastreated with 1.0 M trimethylphosphine in THF (1.76 mL, 1.76 mmol) andstirred for 30 min. The reaction mixture was diluted with DCM (50 mL)and washed with water (50 mL). The aqueous layer was separated andre-extracted with DCM (50 mL). The combined organic extracts were washedwith saturated sodium bicarbonate solution, dried over anhydrous sodiumsulfate, filtered, and concentrated to a colorless foam. Purification byflash column chromatography (100% DCM to 15% MeOH/DCM) afforded thedesired product (0.37 g, 86%) as a white foam. LCMS calculated forC₁₄H₁₉ClFN₂O₂ (M+H)⁺: m/z=301.1; found: 301.0.

Step 3.4-{3-Chloro-6-ethoxy-2-fluoro-5-[1-([1,3]thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl]phenyl}pyrrolidin-2-one

A solution of4-[3-(1-aminoethyl)-5-chloro-2-ethoxy-6-fluorophenyl]pyrrolidin-2-one(30 mg, 0.333 mmol) and 7-chloro[1,3]thiazolo[5,4-d]pyrimidine (25.7 mg,0.15 mmol) [AK-58025, Ark Pharm] in 1-butanol (6.6 mL) was treated withN,N-diisopropylethylamine (0.052 mL, 0.30 mmol) and heated at 114° C.for 2.5 h. The reaction mixture was then concentrated to a yellowresidue. Purification by preparative LCMS (pH 2) afforded the titleproduct (0.033 g, 60%) as a white solid as a mixture of fourdiastereoisomers. ¹H NMR (300 MHz, DMSO-d₆) δ 9.30 (s, 1H), 8.78 (d,J=8.4 Hz, 1H), 8.37 (d, J=1.8 Hz, 1H), 7.82 (s, 1H), 7.71 (d, J=8.3 Hz,1H), 5.91-5.70 (m, 1H), 4.29-4.14 (m, 1H), 4.13-3.96 (m, 1H), 3.95-3.81(m, 1H), 3.60 (dd, J=9.3, 9.3 Hz, 1H), 3.26 (dd, J=7.8, 7.3 Hz, 1H),2.42-2.21 (m, 2H), 1.59-1.32 (m, 6H); LCMS calculated for C₁₉H₂₀ClFN₅O₂S(M+H)⁺: m/z=436.1; found: 435.9.

Example 1A, 1B, 1C, and 1D4-{3-chloro-6-ethoxy-2-fluoro-5-[1-([1,3]thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl]phenyl}pyrrolidin-2-one(Four Diastereoisomers)

The mixture of diastereoisomers of4-{3-chloro-6-ethoxy-2-fluoro-5-[1-([1,3]thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl]phenyl}pyrrolidin-2-onefrom Example 1 were separated by chiral HPLC (Phenomenex Lux CelluloseC-4, 5 micron, 21.2×250 mm, 35% ethanol in hexanes, 18 mL/min) to affordthe individual diastereoisomers.

Example 1A. (Peak 1): ¹H NMR (300 MHz, DMSO-d₆) δ 9.29 (s, 1H), 8.75 (d,J=8.4 Hz, 1H), 8.36 (s, 1H), 7.82 (s, 1H), 7.71 (d, J=8.6 Hz, 1H),5.88-5.70 (m, 1H), 4.30-4.13 (m, 1H), 4.12-3.96 (m, 1H), 3.95-3.82 (m,1H), 3.58 (dd, J=9.4, 9.4 Hz, 1H), 3.30-3.19 (m, 1H), 2.64-2.52 (m, 1H),2.32 (dd, J=18.2, 8.9 Hz, 1H), 1.57-1.34 (m, 6H); LCMS calculated forC₁₉H₂₀ClFN₅O₂S (M+H)⁺: m/z=436.1; found: 435.9.

Example 1B. (Peak 2): ¹H NMR (300 MHz, DMSO-d₆) δ 9.29 (s, 1H), 8.76 (d,J=8.6 Hz, 1H), 8.36 (s, 1H), 7.82 (s, 1H), 7.72 (d, J=8.6 Hz, 1H),5.90-5.70 (m, 1H), 4.32-4.13 (m, 1H), 4.12-3.95 (m, 1H), 3.94-3.79 (m,1H), 3.62 (dd, J=9.0, 9.0 Hz, 2H), 3.33-3.21 (m, 1H), 2.65-2.52 (m, 1H),2.29 (dd, J=17.3, 7.9 Hz, 1H), 1.61-1.37 (m, 6H); LCMS calculated forC₁₉H₂₀ClFN₅O₂S (M+H)⁺: m/z=436.1; found: 435.9.

Example 1C. (Peak 3): ¹H NMR (300 MHz, DMSO-d₆) δ 9.29 (s, 1H), 8.76 (d,J=8.5 Hz, 1H), 8.36 (s, 1H), 7.82 (s, 1H), 7.72 (d, J=8.6 Hz, 1H),5.89-5.71 (m, 1H), 4.30-4.15 (m, 1H), 4.12-3.95 (m, 1H), 3.93-3.79 (m,1H), 3.62 (dd, J=9.6, 9.6 Hz, 1H), 3.30-3.21 (m, 1H), 2.63-2.51 (m, 1H),2.29 (dd, J=16.5, 7.3 Hz, 1H), 1.58-1.35 (m, 6H); LCMS calculated forC₁₉H₂₀ClFN₅O₂S (M+H)⁺: m/z=436.1; found: 435.9.

Example 1D. (Peak 4): ¹H NMR (300 MHz, DMSO-d₆) δ 9.29 (s, 1H), 8.75 (d,J=8.5 Hz, 1H), 8.36 (s, 1H), 7.82 (s, 1H), 7.71 (d, J=8.6 Hz, 1H),5.94-5.64 (m, 1H), 4.28-4.15 (m, 1H), 4.11-3.96 (m, 1H), 3.95-3.82 (m,1H), 3.58 (dd, J=9.8, 9.8 Hz, 1H), 3.30-3.20 (m, 1H), 2.67-2.53 (m, 1H),2.32 (dd, J=18.4, 8.9 Hz, 1H), 1.58-1.37 (m, 6H); LCMS calculated forC₁₉H₂₀ClFN₅O₂S (M+H)⁺: m/z=436.1; found: 435.9.

Example 24-{3-Chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-one

A suspension of4-[3-(1-aminoethyl)-5-chloro-2-ethoxy-6-fluorophenyl]pyrrolidin-2-one(93.0 mg, 0.309 mmol) and 4-chloropyrido[3,2-d]pyrimidine (76.8 mg,0.464 mmol) [093654, Oakwood Chemical] in 1-butanol (6.2 mL) was treatedwith N,N-diisopropylethylamine (0.162 mL, 0.928 mmol) and heated at 105°C. for 30 min. The reaction mixture was then concentrated to a yellowresidue. Purification by flash column chromatography (100% DCM to 10%MeOH/DCM) afforded the crude product. Further purification bypreparative LCMS (pH 10) afforded the title product (0.101 g, 76%) as awhite solid as a mixture of four diastereoisomers. ¹H NMR (300 MHz,DMSO-d₆) δ 8.93 (d, J=8.2 Hz, 1H), 8.85 (dd, J=4.2, 1.5 Hz, 1H), 8.45(d, J=1.5 Hz, 1H), 8.16-8.07 (m, 1H), 7.90-7.73 (m, 3H), 5.90-5.69 (m,1H), 4.32-4.15 (m, 1H), 4.12-3.96 (m, 1H), 3.96-3.80 (m, 1H), 3.60 (dd,J=10.7, 10.7 Hz, 1H), 3.29-3.21 (m, 1H), 2.63-2.55 (m, 1H), 2.39-2.21(m, 1H), 1.52 (d, J=6.8 Hz, 3H), 1.45 (t, J=6.3 Hz, 3H); LCMS calculatedfor C₂₁H₂₂ClFN₅O₂ (M+H)⁺: m/z=430.1; found: 429.9.

Example 2A, 2B, 2C, and 2D4-{3-chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-one(Four Diastereoisomers)

The mixture of diastereoisomers of4-{3-chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-onewere separated by chiral HPLC (Phenomenex Lux Cellulose C-4, 5 micron,21.2×250 mm, 30% ethanol in hexanes, 18 mL/min) to afford thediastereoisomers of Examples 2A and 2B. The diastereoisomers of Examples2C and 2D eluted together and were further purified by chiral HPLC(CHIRALPAK IA, 5 micron, 20×250 mm, 30% ethanol in hexanes, 12 mL/min)to yield individual diastereoisomers.

Example 2A (Peak 1): ¹H NMR (300 MHz, DMSO-d₆) δ 8.93 (d, J=8.5 Hz, 1H),8.85 (dd, J=4.2, 1.5 Hz, 1H), 8.45 (s, 1H), 8.11 (dd, J=8.5, 1.5 Hz,1H), 7.90-7.74 (m, 3H), 5.87-5.74 (m, 1H), 4.30-4.16 (m, 1H), 4.12-3.96(m, 1H), 3.96-3.82 (m, 1H), 3.58 (dd, J=9.6, 9.6 Hz, 1H), 3.30-3.18 (m,1H), 2.65-2.54 (m, 1H), 2.33 (dd, J=17.4, 9.6 Hz, 1H), 1.52 (d, J=6.9Hz, 3H), 1.45 (t, J=6.9 Hz, 3H); LCMS calculated for C₂₁H₂₂ClFN₅O₂(M+H)⁺: m/z=430.1; found: 430.0.

Example 2B (Peak 2): ¹H NMR (300 MHz, DMSO-d₆) δ 8.93 (d, J=8.6 Hz, 1H),8.85 (dd, J=4.2, 1.5 Hz, 1H), 8.44 (s, 1H), 8.11 (dd, J=8.5, 1.5 Hz,1H), 7.90-7.73 (m, 3H), 5.88-5.75 (m, 1H), 4.32-4.16 (m, 1H), 4.13-3.96(m, 1H), 3.94-3.80 (m, 1H), 3.62 (dd, J=9.2, 9.2 Hz, 1H), 3.31-3.24 (m,1H), 2.62-2.52 (m, 1H), 2.30 (dd, J=17.0, 8.6 Hz, 1H), 1.52 (d, J=6.9Hz, 3H), 1.45 (t, J=6.9 Hz, 3H); LCMS calculated for C₂₁H₂₂ClFN₅O₂(M+H)⁺: m/z=430.1; found: 429.9.

Example 2C (Peak 3): ¹H NMR (300 MHz, DMSO-d₆) δ 8.93 (d, J=8.6 Hz, 1H),8.85 (dd, J=4.2, 1.5 Hz, 1H), 8.44 (s, 1H), 8.11 (dd, J=8.5, 1.5 Hz,1H), 7.89-7.74 (m, 3H), 5.87-5.75 (m, 1H), 4.33-4.16 (m, 1H), 4.13-3.96(m, 1H), 3.94-3.80 (m, 1H), 3.62 (dd, J=9.5, 9.5 Hz, 1H), 3.30-3.23 (m,1H), 2.61-2.52 (m, 1H), 2.30 (dd, J=16.7, 8.7 Hz, 1H), 1.53 (d, J=6.9Hz, 3H), 1.45 (t, J=6.9 Hz, 3H); LCMS calculated for C₂₁H₂₂ClFN₅O₂(M+H)⁺: m/z=430.1; found: 429.9.

Example 2D (Peak 4): ¹H NMR (300 MHz, DMSO-d₆) δ 8.93 (d, J=8.6 Hz, 1H),8.85 (dd, J=4.2, 1.5 Hz, 1H), 8.45 (s, 1H), 8.11 (dd, J=8.5, 1.5 Hz,1H), 7.88-7.74 (m, 3H), 5.87-5.74 (m, 1H), 4.30-4.15 (m, 1H), 4.12-3.96(m, 1H), 3.96-3.83 (m, 1H), 3.59 (dd, J=9.2, 9.2 Hz, 1H), 3.31-3.21 (m,1H), 2.65-2.54 (m, 1H), 2.33 (dd, J=16.6, 8.8 Hz, 1H), 1.52 (d, J=6.9Hz, 3H), 1.45 (t, J=6.9 Hz, 3H); LCMS calculated for C₂₁H₂₂ClFN₅O₂(M+H)⁺: m/z=430.1; found: 429.9.

Example 35-{3-Chloro-6-methoxy-2-methyl-5-[(1S)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}-N,N-dimethylpyridine-2-carboxamidebis(trifluoroacetate)

Step 1.tert-Butyl{(1S)-1-[5-chloro-3-(6-cyanopyridin-3-yl)-2-methoxy-4-methylphenyl]ethyl}carbamate

A mixture oftert-butyl[(1S)-1-(3-bromo-5-chloro-2-methoxy-4-methylphenyl)ethyl]carbamate(0.31 g, 0.82 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile(0.24 g, 1.1 mmol), and[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II),complex withdichloromethane (1:1) (0.13 g, 0.16 mmol) in acetonitrile (3.7 mL) waspurged with N₂ and then stirred at 95° C. for 2 h. The mixture was thendiluted with ethyl acetate and washed with water. The organic layer wasdried over MgSO₄, concentrated, and purified by silica gel (eluting with0-30% EtOAc/hexanes) to afford the desired product (0.3 g, 91%). LCMScalculated for C₂₁H₂₅ClN₃O₃ (M+H)⁺: m/z=402.2; found: 402.1.

Step 2.5-(3-{(1S)-1-[(tert-Butoxycarbonyl)amino]ethyl}-5-chloro-2-methoxy-6-methylphenyl)pyridine-2-carboxylicacid

A mixture oftert-butyl{(1S)-1-[5-chloro-3-(6-cyanopyridin-3-yl)-2-methoxy-4-methylphenyl]ethyl}carbamate(0.30 g, 0.75 mmol) and 1.0 M sodium hydroxide in water (3.7 mL, 3.7mmol) in ethanol (3.7 mL) was heated at 90° C. for 6 h in a sealed tube.The mixture was then concentrated to remove ethanol. The resultingresidue was cooled to 0° C., acidified with 1N HCl to pH=3, and thenextracted with ethyl acetate. The combined extracts were dried overMgSO₄ and concentrated to afford the desired product (0.27 g, 86%). LCMScalculated for C₂₁H₂₆ClN₂O₅ (M+H)⁺: m/z=421.2; found: 421.1.

Step 3.tert-Butyl[(1S)-1-(5-chloro-3-{6-[(dimethylamino)carbonyl]pyridin-3-yl}-2-methoxy-4-methylphenyl)ethyl]carbamate

To a solution of5-(3-{(1S)-1-[(tert-butoxycarbonyl)amino]ethyl}-5-chloro-2-methoxy-6-methylphenyl)pyridine-2-carboxylicacid (80 mg, 0.2 mmol), dimethylamine hydrochloride (18 mg, 0.22 mmol)and benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (100 mg, 0.23 mmol) in N,N-dimethylformamide (1 mL)was added N,N-diisopropylethylamine (0.099 mL, 0.57 mmol). The mixturewas stirred at room temperature for 2 h then diluted with water. Theresulting mixture was extracted with ethyl acetate and the combinedorganic layers were dried over MgSO₄, filtered, and concentrated. Theresulting residue was purified on silica gel (eluting with 0-40%EtOAc/dichloromethane) to afford the desired product.

Step 4.5-{3-[(1S)-1-Aminoethyl]-5-chloro-2-methoxy-6-methylphenyl}-N,N-dimethylpyridine-2-carboxamidedihydrochloride

A mixture of tert-butyl[(1S)-1-(5-chloro-3-{6-[(dimethylamino)carbonyl]pyridin-3-yl}-2-methoxy-4-methylphenyl)ethyl]carbamate(0.34 g, 0.76 mmol) in methylene chloride (0.5 mL) was treated with 4.0M hydrogen chloride in dioxane (1.5 mL) at room temperature for 2 h andthen concentrated to dryness to afford the desired product which wasused directly in the next step. LCMS calculated for C₁₈H₂₃ClN₃O₂ (M+H)⁺:m/z=348.1; found: 348.1.

Step 5.5-{3-Chloro-6-methoxy-2-methyl-5-[(1S)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}-N,N-dimethylpyridine-2-carboxamidebis(trifluoroacetate)

A mixture of5-{3-[(1S)-1-aminoethyl]-5-chloro-2-methoxy-6-methylphenyl}-N,N-dimethylpyridine-2-carboxamidedihydrochloride (17 mg, 0.040 mmol), 4-chloropyrido[3,2-d]pyrimidine(6.7 mg, 0.040 mmol) and N,N-diisopropylethylamine (35 μL, 0.20 mmol) in1-butanol (0.4 mL) was heated at 120° C. for 2 h. The mixture waspurified on prep-LCMS (Sunfire Prep C₁₈ ₅ μm 30×10 mm column, flow rate60 mL/min, eluting with a gradient of acetonitrile and water with 0.05%TFA) to afford the title product (2.7 mg, 14%). LCMS calculated forC₂₅H₂₆ClN₆O₂ (M+H)⁺: m/z=477.2; found: 477.2.

Example 4.4-Chloro-N-ethyl-3′,5′-difluoro-3-methyl-6-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]biphenyl-2-carboxamide

Step 1. 3-Acetyl-5-chloro-2-hydroxy-6-methylbenzonitrile

A mixture of 1-(3-bromo-5-chloro-2-hydroxy-4-methylphenyl)ethanone (4.85g, 18.4 mmol) and copper cyanide (2.47 g, 27.6 mmol) inN-methylpyrrolidinone (15 mL) was heated at 200° C. for 1 h. Aftercooling to room temperature, the mixture was diluted with ethyl acetateand 1 N HCl. The layers were separated and the aqueous layer wasextracted with ethyl acetate. The combined organic layers were washedwith water, brine, dried over magnesium sulfate, then filtered andconcentrated to dry under reduced pressure. The residue (3.7 g, 96%) wasused directly in the next step without further purification. LCMScalculated for C₁₀H₉ClNO₂ (M+H)⁺: m/z=210.0; found: 210.1.

Step 2. 6-Acetyl-4-chloro-2-cyano-3-methylphenyltrifluoromethanesulfonate

To a mixture of 3-acetyl-5-chloro-2-hydroxy-6-methylbenzonitrile (3.70g, 17.6 mmol) in methylene chloride (70 mL) was added triethylamine (7.4mL, 53 mmol) followed by trifluoromethanesulfonic anhydride (4.4 mL, 26mmol) at −78° C. The reaction mixture was gradually warmed to roomtemperature and stirred at room temperature for 30 min. After quenchingwith water, the mixture was extracted with dichloromethane. The combinedorganic layers were washed with brine, dried over sodium sulfate,filtered, and concentrated to dryness. The residue was purified onsilica gel (eluting with 0 to 40% EtOAc in hexanes) to afford thedesired product (2.54 g, 42%). LCMS calculated for C₁₁H₈ClF₃NO₄S (M+H)⁺:m/z=342.0; found: 342.1.

Step 3. 6-Acetyl-4-chloro-3′,5′-difluoro-3-methylbiphenyl-2-carbonitrile

A biphasic solution of 6-acetyl-4-chloro-2-cyano-3-methylphenyltrifluoromethanesulfonate (4.51 g, 13.2 mmol) and(3,5-difluorophenyl)boronic acid (2.50 g, 15.8 mmol) in toluene (50mL)/0.8 M sodium hydrogencarbonate in water (50 mL, 40 mmol) wasdegassed with N₂. Tetrakis(triphenylphosphine)palladium(0) (0.609 g,0.527 mmol) was then added. The mixture was bubbled with N₂ for 5 minand then heated at 80° C. for 2 h. After cooling to room temperature,the mixture was diluted with ethyl acetate. The layers were separatedand the aqueous layer was extracted with ethyl acetate. The combinedextracts were washed with brine, dried over Na₂SO₄, filtered, andconcentrated to a crude dark solid. This material was then dissolved inCHCl₃ and purified on a silica gel column (eluting with 0-30% ethylacetate/hexanes) to afford the desired product (1.94 g, 48%). LCMScalculated for C₁₆H₁₁ClF₂NO (M+H)⁺: m/z=306.0; found: 306.0.

Step 4.4-Chloro-3′,5′-difluoro-6-(1-hydroxyethyl)-3-methylbiphenyl-2-carbaldehyde

To a mixture of6-acetyl-4-chloro-3′,5′-difluoro-3-methylbiphenyl-2-carbonitrile (2.43g, 7.95 mmol) in methylene chloride (50 mL) was added 1.0 Mdiisobutylaluminum hydride in hexane (19.9 mL, 19.9 mmol) at −78° C. Thereaction mixture was warmed to room temperature over 2 h with stirring.5.0 M Hydrogen chloride in water (70 mL, 400 mmol) was then added slowlyand stirring was continued for 1 h. The resultant mixture was extractedwith ethyl acetate. The combined organic layers were washed with brine,dried over sodium sulfate, filtered, and concentrated to dryness. Theresidue was purified on silica gel (eluting with 0 to 50% ethyl acetatein hexanes) to afford the desired product (2.4 g, 97%). LCMS calculatedfor C₁₆H₁₂ClF₂O (M−OH)⁺: m/z=293.1; found: 293.1.

Step 5.4-Chloro-3′,5′-difluoro-6-(1-hydroxyethyl)-3-methylbiphenyl-2-carboxylicacid

To a solution of4-chloro-3′,5′-difluoro-6-(1-hydroxyethyl)-3-methylbiphenyl-2-carbaldehyde(1.00 g, 3.22 mmol) in methanol (40 mL) was added 1.0 M sodium hydroxidein water (16 mL, 16 mmol), followed by urea hydrogen peroxide (CAS#124-43-6 purchased from Sigma-Aldrich, 1.00 g, 10.6 mmol). Afterstirring overnight at room temperature, the mixture was slowly acidifiedto pH 5 with 1 N HCl then extracted with ethyl acetate. The combinedorganic layers were washed with brine, dried over magnesium sulfate,filtered, and concentrated to dryness under reduced pressure. The cruderesidue (1.05 g, 99.8%) was used directly in the next step.

Step 6.4-Chloro-N-ethyl-3′,5′-difluoro-6-(1-hydroxyethyl)-3-methylbiphenyl-2-carboxamide

A mixture of4-chloro-3′,5′-difluoro-6-(1-hydroxyethyl)-3-methylbiphenyl-2-carboxylicacid (250 mg, 0.76 mmol), ethylamine hydrochloride (94 mg, 1.1 mmol) andbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(0.51 g, 1.1 mmol) in N,N-dimethylformamide (4 mL) was stirred at roomtemperature for 10 min. To the resulting mixture was addedN,N-diisopropylethylamine (0.40 mL, 2.3 mmol). After stirring overnightat room temperature, the reaction mixture was quenched with water andextracted with ethyl acetate. The combined organic layers were washedwith water and brine, dried over magnesium sulfate, filtered, andconcentrated to dryness. The residue was purified on silica gel (elutingwith 0 to 80% ethyl acetate in hexanes) to afford the desired product(0.185 g, 68%). LCMS calculated for C₁₈H₁₉ClF₂NO₂ (M+H)⁺: m/z=354.1;found: 354.0.

Step 7.1-{4-Chloro-6-[(ethylamino)carbonyl]-3′,5′-difluoro-5-methylbiphenyl-2-yl}ethylmethanesulfonate

To a mixture of4-chloro-N-ethyl-3′,5′-difluoro-6-(1-hydroxyethyl)-3-methylbiphenyl-2-carboxamide(185 mg, 0.523 mmol) in methylene chloride (3 mL) was addedN,N-diisopropylethylamine (0.18 mL, 1.0 mmol), followed bymethanesulfonyl chloride (0.061 mL, 0.78 mmol). The reaction was stirredat room temperature for 10 min, quenched by pouring over ice water, andthen extracted with dichloromethane. The combined organic layers werewashed with aqueous sodium bicarbonate, dried over magnesium sulfate,filtered, and evaporated to dryness. The resulting residue (0.226 g,100%) was used directly in the next step without further purification.LCMS calculated for C₁₉H₂₁ClF₂NO₄S (M+H)⁺: m/z=432.1; found: 432.1.

Step 8.6-(1-Azidoethyl)-4-chloro-N-ethyl-3′,5′-difluoro-3-methylbiphenyl-2-carboxamide

To a mixture of1-{4-chloro-6-[(ethylamino)carbonyl]-3′,5′-difluoro-5-methylbiphenyl-2-yl}ethylmethanesulfonate (390 mg, 0.90 mmol) in N,N-dimethylformamide (5 mL) wasadded sodium azide (290 mg, 4.5 mmol). The reaction was stirredovernight at room temperature, then quenched with water and extractedwith ethyl acetate. The combined organic layers were washed with waterand brine, dried over magnesium sulfate, filtered, and concentrated todryness. The crude residue (0.34 g, 99%) was used directly in the nextstep without further purification. LCMS calculated for C₁₈H₁₈ClF₂N₄O(M+H)⁺: m/z=379.1; found: 379.0.

Step 9.6-(1-Aminoethyl)-4-chloro-N-ethyl-3′,5′-difluoro-3-methylbiphenyl-2-carboxamide

To a stirred mixture of6-(1-azidoethyl)-4-chloro-N-ethyl-3′,5′-difluoro-3-methylbiphenyl-2-carboxamide(0.34 g, 0.90 mmol) in tetrahydrofuran (6 mL) and water (1 mL) was added1.0 M trimethylphosphine in THF (1.1 mL, 1.1 mmol). The mixture was thenstirred at room temperature for 1 h. After nitrogen was passed throughthe mixture, the reaction mixture was extracted with dichloromethane.The extracts were washed with brine, dried over magnesium sulfate,filtered, and evaporated to dryness. The crude residue (0.135 g, 43%)was used directly in the next step without further purification. LCMScalculated for C₁₈H₂₀ClF₂N₂O (M+H)⁺: m/z=353.1; found: 353.1.

Step 10.4-Chloro-N-ethyl-3′,5′-difluoro-3-methyl-6-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]biphenyl-2-carboxamide

A mixture of6-(1-aminoethyl)-4-chloro-N-ethyl-3′,5′-difluoro-3-methylbiphenyl-2-carboxamide(30 mg, 0.08 mmol), 4-chloropyrido[3,2-d]pyrimidine (30 mg, 0.2 mmol)and N,N-diisopropylethylamine (0.04 mL, 0.2 mmol) in 1-butanol (0.5 mL)was heated at 100° C. overnight. After evaporating to dryness, theresidue was purified on RP-HPLC (Sunfire Prep C_(18 5 μ)m 30×10 mmcolumn, flow rate 60 mL/min, eluting with a gradient of MeCN and waterwith 0.1% ammonium hydroxide) to afford the desired product. LCMScalculated for C₂₅H₂₃ClF₂N₅O (M+H)⁺: m/z=482.2; found: 482.1.

Example J1((2R,5S)-5-{2-[(1R)-1-Hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile

Step 1. tert-Butyl(4S)-2,2-dimethyl-4-vinyl-1,3-oxazolidine-3-carboxylate

To a suspension of methyl triphenylphosphonium bromide (5.63 g, 15.8mmol) in tetrahydrofuran (140 mL) was added 2.5 M n-butyllithium inhexane (7.35 mL, 18.4 mmol). The deep red solution was stirred at 0° C.for 1 h. Then a solution of tert-butyl(4R)-4-formyl-2,2-dimethyl-1,3-oxazolidine-3-carboxylate (from Aldrich,3.01 g, 13.1 mmol) in tetrahydrofuran (7.3 mL) was added drop wise at 0°C. The red solution was warmed to room temperature and stirred for 12 h.Hexanes was added to the reaction mixture in 4:1 (v/v) ratio. Thesuspension was filtered through Celite and the filtrate concentrated.The resultant residue was purified by flash chromatography (eluting with10% ethyl acetate in hexanes) to give the desired compound as colorlessoil (1.92 g, 64%).

Step 2. tert-Butyl [(1S)-1-(hydroxymethyl)prop-2-en-1-yl]carbamate

To a solution of tert-butyl(4S)-2,2-dimethyl-4-vinyl-1,3-oxazolidine-3-carboxylate (1.90 g, 8.36mmol) in methanol (83 mL) was added p-toluenesulfonic acid monohydrate(0.80 g, 4.2 mmol) at 0° C. The mixture was slowly warmed to roomtemperature overnight. The reaction mixture was diluted with saturatedNaHCO₃ solution, concentrated, and then diluted with ethyl acetate. Theorganic layer was washed with sat. NaHCO₃ (2×) and brine, dried overNa₂SO₄, filtered and concentrated to give the desired product ascolorless oil (1.187 g, 76%). ¹H NMR (400 MHz, CDCl₃) δ 5.81 (1H, m),5.25 (2H, m), 4.90 (1H, m), 4.25 (1H, br s), 3.67 (2H, m), 1.45 (9H, s)ppm.

Step 3. tert-Butyl[(1S)-1-({[1-(hydroxymethyl)prop-2-en-1-yl]oxy}methyl)prop-2-en-1-yl]carbamate

To a flask was charged with tert-butyl[(1S)-1-(hydroxymethyl)prop-2-en-1-yl]carbamate (0.401 g, 2.14 mmol),tris(dibenzylideneacetone)dipalladium(0) (59 mg, 0.064 mmol),N,N′-(1S,2S)-cyclohexane-1,2-diylbis[2-(diphenylphosphino)-1-naphthamide](150 mg, 0.19 mmol), and 4-dimethylaminopyridine (78 mg, 0.64 mmol). Thereaction mixture was purged with N₂ three times, and then methylenechloride (21.3 mL), and 1.0 M triethylborane in THF (130 μL, 0.13 mmol)was added sequentially. After stirring for 10 min, 2-vinyloxirane (0.150g, 2.14 mmol) was added and the resulting mixture was stirred overnight.The reaction was diluted with dichloromethane and sat. NaHCO₃ solution.The organic layer was separated and dried over Na₂SO₄, filtered andconcentrated. The crude residue was purified with flash chromatography(eluting with 0-50% ethyl acetate/hexanes) to give the desired product(0.271 g, 49%). ¹H NMR (300 MHz, CDCl₃) δ 5.85 (1H, m), 5.67 (1H, m),5.84˜5.17 (4H, m), 4.83 (1H, m), 4.30 (1H, br s), 3.83 (1H, m), 3.69(1H, dd, J=4.5 and 6.9 Hz), 3.54 (2H, m), 3.36 (1H, dd, J=4.5 and 6.9Hz), 1.45 (9H, s) ppm.

Step 4.2-({(2S)-2-[(tert-Butoxycarbonyl)amino]but-3-en-1-yl}oxy)but-3-en-1-ylacetate

To a mixture of tert-butyl[(1S)-1-({[1-(hydroxymethyl)prop-2-en-1-yl]oxy}methyl)prop-2-en-1-yl]carbamate(268 mg, 1.04 mmol) in methylene chloride (10 mL) was added withtriethylamine (435 μL, 3.12 mmol). The mixture was cooled to 0° C., andacetyl chloride (150 μL, 2.1 mmol) was added drop wise. The reaction wasstirred at room temperature for 2 h, then quenched with water. Theorganic layer was concentrated and the resultant residue purified onsilica gel (eluting with 20% ethyl acetate/hexanes) to give the desiredproduct (0.26 g, 85%). LCMS calculated for C₁₀H₁₈NO₃ (M−100+H)⁺:m/z=200.1; Found: 200.1.

Step 5.{(5S)-5-[(tert-Butoxycarbonyl)amino]-5,6-dihydro-2H-pyran-2-yl}methylacetate

To a 500 mL 2-neck round bottom flask,benzylidene(dichloro)(1,3-dimesitylimidazolidin-2-id-2-yl)(tricyclohexylphosphoranyl)ruthenium(38 mg, 0.044 mmol) was added. After purged with nitrogen for 3 times,dichloromethane (anhydrous, 8 mL) was added followed by2-({(2S)-2-[(tert-butoxycarbonyl)amino]but-3-en-1-yl}oxy)but-3-en-1-ylacetate (265 mg, 0.885 mmol). The reaction mixture was stirred at roomtemperature for 15 h. The mixture was concentrated in vacuo. The residuewas purified via flash chromatography (eluting with hexanes to 25% EtOAcin hexanes) to give the desired product as a brown oil (0.205 g, 85%).LCMS calculated for C₉H₁₄NO₅ (M+H−Bu+H)⁺: m/z=216.1; Found: 216.1. ¹HNMR (300 MHz, CDCl₃) δ 5.94 (0.17H, m), 5.84 (0.83H, m), 5.69 (1H, m),4.89 (0.13H, m), 4.70 (0.83H, m), 4.25 (1H, m), 4.05 (4H, m), 3.56(0.13H, m), 3.38 (0.87H, m), 2.04 (2.49H, s), 2.03 (0.51H, m), 1.38 (9H,s) ppm (The product was a ˜5:1 mixture of trans- and cis-isomers).

Step 6. [(5S)-5-Amino-5,6-dihydro-2H-pyran-2-yl]methyl acetate

To a solution of{(5S)-5-[(tert-butoxycarbonyl)amino]-5,6-dihydro-2H-pyran-2-yl}methylacetate (205 mg, 0.756 mmol) in methylene chloride (5.2 mL) was added4.0 M hydrogen chloride in dioxane (1.5 mL, 6.0 mmol). The reactionsolution was stirred at room temperature for 6 h. The solvent wasremoved under reduced pressure to give the desired product as whitesolid. LCMS calculated for C₈H₁₄NO₃ (M+H)⁺: m/z=172.1; Found: 172.1.

Step 7.{(5S)-5-[(6-Nitrothieno[3,2-b]pyridin-7-yl)amino]-5,6-dihydro-2H-pyran-2-yl}methylacetate

A mixture of 7-chloro-6-nitrothieno[3,2-b]pyridine (156 mg, 0.727 mmol),[(5S)-5-amino-5,6-dihydro-2H-pyran-2-yl]methyl acetate (129 mg, 0.754mmol) and N,N-diisopropylethylamine (0.26 mL, 1.5 mmol) in isopropylalcohol (1.7 mL) was heated at 90° C. for 2 h. The reaction mixture wasconcentrated and purified with flash chromatography to give the desiredproduct (0.21 g 83%). LCMS calculated for C₁₅H₁₆N₃O₅S (M+H)⁺: m/z=350.1;Found: 350.0.

Step 8.{(5S)-5-[(6-Aminothieno[3,2-b]pyridin-7-yl)amino]tetrahydro-2H-pyran-2-yl}methylacetate

A mixture of{(5S)-5-[(6-nitrothieno[3,2-b]pyridin-7-yl)amino]-5,6-dihydro-2H-pyran-2-yl}methylacetate (210 mg, 0.600 mmol) and 10% palladium on carbon (0.21 g) inmethanol (4.0 mL) was subjected to balloon pressure of H₂ at roomtemperature for 2 h. The mixture was filtered, and the filtrate wasconcentrated and purified with flash chromatography (eluting with 15%methanol in dichloromethane) to give the desired product (145 mg, 75%).LCMS calculated for C₁₅H₂₀N₃O₃S (M+H)⁺: m/z=322.1; Found: 322.0.

Step 9.(1R)-1-{1-[(3S)-6-(Hydroxymethyl)tetrahydro-2H-pyran-3-yl]-H-imidazo[4,5-d]thieno[3,2b]pyridin-2-yl}ethanol

A mixture of (2R)-2-hydroxypropanamide (131 mg, 1.47 mmol) andtriethyloxonium tetrafluoroborate (263 mg, 1.38 mmol) in THF (2 mL) wasstirred at room temperature for 2 h. The solvent was removed and theresidue dissolved in ethanol (0.85 mL) and added to a suspension of{(5S)-5-[(6-aminothieno[3,2-1-b]pyridin-7-yl)amino]tetrahydro-2H-pyran-2-yl}methylacetate (145 mg, 0.451 mmol) in ethanol (3.1 mL). The mixture wasstirred at 80° C. for 1 h. The reaction was cooled to room temperatureand diluted with water (1.0 mL). Lithium hydroxide (32.4 mg, 1.35 mmol)was added, and the mixture was stirred for 2 h. The reaction mixture wasdiluted with methanol and purified with prep-LCMS (XBridge C₁₈ column,eluting with a gradient of acetonitrile/water containing 0.1% ammoniumhydroxide, at flow rate of 60 mL/min) to give the desired product aswhite solid (95 mg, 63%). LCMS calculated for C₁₆H₂₀N₃O₃S (M+H)⁺:m/z=334.1; Found: 334.0.

Step 10:((2R,5S)-5-{2-[(1R)-1-Hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)methyl4-methylbenzenesulfonate and((2S,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)methyl4-methylbenzenesulfonate

To a solution of(1R)-1-{1-[(3S)-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-2-yl}ethanol(100 mg, 0.300 mmol) (previous step) in methylene chloride (3.4 mL) andpyridine (0.146 mL, 1.80 mmol) was added p-toluenesulfonyl chloride(57.2 mg, 0.300 mmol) and 4-dimethylaminopyridine (1.8 mg, 0.015 mmol)at 0° C. The reaction mixture was allowed to warm to room temperatureovernight. The reaction mixture was concentrated, diluted with methanol,and purified with prep-LCMS (XBridge C₁₈ column, eluting with a gradientof acetonitrile/water containing 0.1% ammonium hydroxide, at flow rateof 60 mL/min) to give two peaks. On analytic HPLC (Waters SunFire C18,2.1×50 mm, 5 μM; Flow rate 3 mL/min; Injection volume 2 μL; At gradientfrom 2 to 80% B in 3 minutes (A=water with 0.025% TFA, B=acetonitrile)):First peak (45.3 mg, 31%) retention time 1.81 min, LCMS calculated forC₂₃H₂₆N₃O₅S₂ (M+H)⁺: m/z=488.1; Found: 488.1. Second peak (8.5 mg, 5.8%)retention time 1.88 min, LCMS calculated for C₂₃H₂₆N₃O₅S₂ (M+H)⁺:m/z=488.1; Found: 488.1.

Step 11.((2R,5S)-5-{2-[(1R)-1-Hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrde

A mixture of((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)methyl4-methylbenzenesulfonate (from 1st peak of previous step, 27 mg, 0.055mmol) and sodium cyanide (4.5 mg, 0.092 mmol) in dimethyl sulfoxide (0.4mL) was stirred at 50° C. for 4 h. After cooling, the mixture wasdiluted with methanol and purified with prep-LCMS (XBridge C₁₈ column,eluting with a gradient of acetonitrile/water containing 0.1% ammoniumhydroxide, at flow rate of 30 mL/min) to give the desired product (14.5mg, 76%). LCMS calculated for C₁₇H₁₉N₄O₂S (M+H )⁺: m/z=343.1; Found:343.0. ¹H NMR (DMSO-d₆, 500 MHz) δ 9.51 (1H, s), 8.45 (1H, d, J=5.5 Hz),7.97 (1H, d, J=5.5 Hz), 5.31 (1H, m), 5.20 (1H, m), 4.31 (1H, m), 4.23(1H, m), 4.02 (1H, m), 2.96 (1H, dd, J=17.0 and 4.5 Hz), 2.85 (1H, dd,J=17.0 and 4.5 Hz), 2.66 (1H, m), 2.26 (1H, m), 2.09 (1H, m), 1.73 (1H,m), 1.69 (3H, d, J=6.5 Hz) ppm.

Example J1a.((2R,5S)-5-{2-[(1R)-1-Hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrilehydrate

((2R,5S)-5-{2-[(1R)-1-Hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile(52 mg, 0.15 mmol) from Example 25 was crystallized from a mixture ofacetonitrile (8 mL) and water (4 mL). The resulting colorless prismcrystal collected was suitable for X-ray crystal structure analysis.

Crystal data shows: ˜0.520×0.180×0.100 mm, orthorhombic, P212121,a=6.962(3) Å, b=11.531(4) Å, c=20.799(7) Å, Vol=1669.6(10) Å³, Z=4,T=−100° C., Formula weight=359.42, Density=1.430 g/cm³, μ(Mo)=0.22 mm⁻¹.

Data collection was done on a Bruker SMART APEX-II CCD system, MoKalpharadiation, standard focus tube, anode power=50 kV×42 mA, crystal toplate distance=5.0 cm, 512×512 pixels/frame, beam center=(256.13,253.14), total frames=1151, oscillation/frame=0.50°, exposure/frame=10.1sec/frame, SAINT integration, hkl min/max=(−9, 9, −15, 15, −27, 27),data input to shelx=17025, unique data=3975, two-theta range=3.92 to55.72°, completeness to two-theta 55.72=99.80%, R(int-xl)=0.0681, SADABScorrection applied.

Structure was solved using XS(Shelxtl), refined using shelxtl softwarepackage, refinement by full-matrix least squares on F², scatteringfactors from Int. Tab. Vol C Tables 4.2.6.8 and 6.1.1.4, number ofdata=3975, number of restraints=0, number of parameters=235,data/parameter ratio=16.91, goodness-of-fit on F²=1.04, Rindices[I>4sigma(I)] R1=0.0505, wR2=0.1242, R indices(all data)R1=0.0769, wR2=0.1401, max difference peak and hole=0.724 and −0.277e/Å³, refined flack parameter=−0.12(13), All of the CH hydrogen atomswere refined using a riding model. The OH hydrogens were found from adifference map and fully refined.

Results showed that the asymmetric unit contains one molecule and onewater as shown with thermal ellipsoids drawn to the 50% probabilitylevel. The stereochemistry at each of three stereocenters (as indicatedin the name and structure of the compound above) was confirmed. Theflack parameter refined to 0.28(24) indicating the correct enantiomericsetting.

Example J2 4-[3-(Cyanomethyl)-3-(3′,5′-dimethyl-1H,PH-4,4′-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide

Step 1: 2,4,5-Trifluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide

To a solution of 2,4,5-trifluorobenzoic acid (5.00 g, 28.4 mmol) inacetonitrile (50 mL) was added N,N-dimethylformamide (40 μL) followed byaddition of oxalyl chloride (3.60 mL, 42.6 mmol). After 90 min, thevolatiles were removed under reduced pressure. The residue wasco-evaporated with acetonitrile (50 mL). The residue was then dissolvedin methylene chloride (50 mL). This solution was added drop-wise into acooled (ice bath) mixture of (2S)-1,1,1-trifluoropropan-2-aminehydrochloride (5.52 g, 36.9 mmol) (from Synquest, 98% ee) in toluene(100 mL) and 0.5 M sodium hydroxide aqueous solution (142 mL, 71.0mmol). After addition, the ice bath was removed, and the reaction wasallowed to warm to rt. The reaction was stirred overnight. The organiclayer was separated. The aqueous layer was extracted with methylenechloride (50 mL). The combined organic layers were washed with 20% brine(75 mL) and water (2×75 mL), dried over MgSO₄, filtered and concentratedunder reduced pressure to afford the desired product (6.49 g, 84%) whichwas directly used in the next step without further purification. ¹H NMR(300 MHz, DMSO-d₆) δ 9.01 (d, J=7.6 Hz, 1H), 7.92-7.50 (m, 2H), 4.76 (m,1H), 1.31 (d, J=7.0 Hz, 3H) ppm. LCMS cacld. for C₁₀H₈F₆NO (M+1)⁺:m/z=272.0; Found: 272.0.

Step 2: 2,5-Difluoro-4-(3-hydroxyazetidin-1-yl)-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide

A mixture of2,4,5-trifluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide (6.39 g,23.6 mmol), azetidin-3-ol hydrochloride (3.19 g, 28.3 mmol) and1,8-diazabicyclo[5.4.0]undec-7-ene (8.81 mL, 58.9 mmol) in acetonitrile(25 mL) was stirred at 80° C. for 2 h. The reaction mixture was dilutedwith EtOAc (75 mL) and washed with 1N HCl (50 mL), 1N NaHCO₃ (60 mL),20% brine (50 mL) and water (75 mL). The aqueous layers were extractedwith EtOAc (100 mL). The organic layers were combined, dried over MgSO₄,filtered and concentrated under reduced pressure to yield the de11siredproduct (7.59 g, 91.8%). ¹H NMR (300 MHz, DMSO-d₆) δ 8.38 (dd, J=8.9,1.9 Hz, 1H), 7.27 (dd, J=12.8, 6.5 Hz, 1H), 6.38 (dd, J=12.3, 7.5 Hz,1H), 5.71 (d, J=6.4 Hz, 1H), 4.74 (dp, J=15.3, 7.6 Hz, 1H), 4.62-4.46(m, 1H), 4.30-4.15 (m, 2H), 3.71 (m, 2H), 1.29 (d, J=7.1 Hz, 3H) ppm.LCMS cacld. for C₁₃H₁₄F₅N₂O₂ (M+1)⁺: m/z=325.1; Found: 325.1.

Step 3: 2,5-Difluoro-4-(3-oxoazetidin-1-yl)-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide

To a solution of2,5-difluoro-4-(3-hydroxyazetidin-1-yl)-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide(7.57 g, 23.3 mmol) in methylene chloride (93 mL) was added iodobenzenediacetate (9.40 g, 29.2 mmol) and 2,2,6,6-tetramethyl-l-piperidinyloxyfree radical (1.82 g, 11.7 mmol) (TEMPO) at room temperature. Thereaction mixture was stirred at room temperature overnight. The mixturewas diluted with EtOAc (100 mL), washed with 0.5N NaHCO₃ (2×80 mL), 20%brine (100 mL) and water (100 mL). The aqueous layers were extractedwith ethyl acetate (75 mL). The organic extracts were combined, driedover MgSO₄, filtered and concentrated under reduced pressure. Theresidue was purified by flash chromatography on a silica gel columneluting with 0% to 5% ethyl acetate in methylene chloride to afford thecrude product which was recrystallized from MTBE (50 mL) and heptane(100 mL) to give the desired product (5.44g, 72%) as colorless solid. ¹HNMR (300 MHz, DMSO-d₆) δ 8.52 (d, J=8.0 Hz, 1H), 7.36 (dd, J=12.5, 6.5Hz, 1H), 6.63 (dd, J=12.1, 7.6 Hz, 1H), 4.90 (d, J=2.1 Hz, 4H),4.86-4.68 (m, 1H), 1.31 (d, J=7.1 Hz, 3H) ppm. LCMS cacld. forC₁₃H₁₂F₅N₂O₂ (M+1)⁺: m/z=323.1; Found: 323.0.

Step 4:4-[3-(Cyanomethylene)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide

Diethyl cyanomethylphosphonate (1.95 mL, 11.8 mmol) was added drop-wiseto a cooled (ice bath) solution of 1.0 M potassium tert-butoxide in THF(11.8 mL, 11.8 mmol) which was diluted with tetrahydrofuran (12 mL). Thebath was removed and the reaction was warmed to room temperature, andstirred for 90 min. The reaction solution was cooled with an ice bathagain. The above prepared solution was then added over 12 min to acooled (ice-bath) solution of2,5-difluoro-4-(3-oxoazetidin-1-yl)-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide(4.00 g, 12.4 mmol) in tetrahydrofuran (50 mL). The reaction mixture wasstirred for 30 min. The ice bath was removed, and the reaction wasstirred at room temperature overnight, then quenched by the addition of20% brine (75 mL) and ethyl acetate (75 mL). The organic layer wasseparated. The aqueous layer was extracted with ethyl acetate (50 mL).The combined organic layers were dried over MgSO₄, filtered andconcentrated under reduced pressure. The residue was purified by flashchromatography on a silica gel column with ethyl acetate in hexanes (0%to 30%) to yield the desired product (2.6 g). ¹H NMR (400 MHz, DMSO-d₆)δ 8.59-8.37 (m, 1H), 7.33 (dd, J=12.5, 6.4 Hz, 1H), 6.59 (dd, J=12.0,7.4 Hz, 1H), 5.88 (m, 1H), 4.94-4.75 (m, 4H), 4.76 (m, 1H), 1.31 (d,J=7.1 Hz, 3H) ppm. LCMS cacld. for C₁₅H₁₃F₅N₃O (M+1)⁺: m/z=346.1; Found:346.1.

Step 5:4-{3-(Cyanomethyl)-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide

A mixture of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(1.00 g, 5.15 mmol),4-[3-(cyanomethylene)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide(1.78 g, 5.15 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (0.31 mL, 2.1mmol) in acetonitrile (20.2 mL) was heated at 50° C. overnight. Aftercooling, the solvent was removed under reduced pressure. The residue wasused in the next step without further purification. LCMS cacld. forC₂₄H₂₈BF₅N₅O₃ (M+1)⁺: m/z=540.2; Found: 540.1.

Step 6:4-[3-(Cyanomethyl)-3-(3′,5′-dimethyl-1H,1′H-4,4′-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide

A mixture of4-{3-(cyanomethyl)-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-y)-1H-pyrazol-1-yl]azetidin-1-yl}-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide(329 mg, 0.610 mmol), 4-bromo-3,5-dimethyl-1H-pyrazole (206 mg, 1.18mmol), tetrakis(triphenylphosphine)palladium(0) (110 mg, 0.098 mmol) andsodium carbonate (320 mg, 3.0 mmol) in 1,4-dioxane (10 mL)/water (5 mL)was purged with nitrogen and stirred at 110° C. for 1 h. The reactionmixture was diluted with EtOAc, washed with water and brine,concentrated. The residue was purified first with silica gel (elutingwith 0-100% EtOAc/hexanes followed by 10% methanol/dichloromethane), andthen by prep-LCMS (XBridge C18 column, eluting with a gradient ofacetonitrile/water containing 0.1% ammonium hydroxide, at flow rate of60 mL/min) to give the desired product (30 mg, 9.7%). ¹H NMR (500 MHz,DMSO-d₆) δ 12.17 (1H, s), 8.45 (1H, d, J=8.0 Hz), 8.10 (1H, s), 7.70(1H, s), 7.34 (1H, m), 6.61 (1H, s), 4.77 (1H, m), 4.62 (2H, d, J=9.0Hz), 4.39 (1H, d, J=9.0 Hz), 3.64 (2H, s), 2.22 (6H, s), 1.31 (6H, d,J=7.0 Hz) ppm. LCMS calculated for C₂₃H₂₃F₅N₇O (M+H)⁺: m/z=508.2; Found:508.0.

Example A1 PI3K Enzyme Assay

PI3-Kinase luminescent assay kit including lipid kinase substrate,D-myo-phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2)D(+)-sn-1,2-di-O-octanoylglyceryl, 3-O-phospho linked (PIP2),biotinylated I(1,3,4,5)P4, PI(3,4,5)P3 Detector Protein is purchasedfrom Echelon Biosciences (Salt Lake City, Utah). AlphaScreen™ GSTDetection Kit including donor and acceptor beads was purchased fromPerkinElmer Life Sciences (Waltham, Mass. PI3Kδ (p110δ/p85α) ispurchased from Millipore (Bedford, Mass.). ATP, MgCl₂, DTT, EDTA, HEPESand CHAPS are purchased from Sigma-Aldrich (St. Louis, Mo.).

AlphaScreen™ Assay for PI3Kδ

The kinase reaction are conducted in 384-well REMP plate from ThermoFisher Scientific in a final volume of 40 μL. Inhibitors are firstdiluted serially in DMSO and added to the plate wells before theaddition of other reaction components. The final concentration of DMSOin the assay is 2%. The PI3K assays are carried out at room temperaturein 50 mM HEPES, pH 7.4, 5 mM MgCl₂, 50 mM NaCl, 5 mM DTT and CHAPS0.04%. Reactions are initiated by the addition of ATP, the finalreaction mixture consisted of 20 μM PIP2, 20 μM ATP, 1.2 nM PI3Kδ areincubated for 20 minutes. 10 μL of reaction mixture are then transferredto 5 μL 50 nM biotinylated I(1,3,4,5)P4 in quench buffer: 50 mM HEPES pH7.4, 150 mM NaCl, 10 mM EDTA, 5 mM DTT, 0.1% Tween-20, followed with theaddition of 10 μL AlphaScreen™ donor and acceptor beads suspended inquench buffer containing 25 nM PI(3,4,5)P3 detector protein. The finalconcentration of both donor and acceptor beads is 20 mg/ml. After platesealing, the plate are incubated in a dark location at room temperaturefor 2 hours. The activity of the product is determined on Fusion-alphamicroplate reader (Perkin-Elmer). IC₅₀ determination is performed byfitting the curve of percent control activity versus the log of theinhibitor concentration using the GraphPad Prism 3.0 software.

Example A2 PI3K Enzyme Assay

Materials: Lipid kinase substrate, phosphoinositol-4,5-bisphosphate(PIP2), are purchased from Echelon Biosciences (Salt Lake City, Utah).PI3K isoforms α, β, δ and γ are purchased from Millipore (Bedford,Mass.). ATP, MgCl₂, DTT, EDTA, MOPS and CHAPS are purchased fromSigma-Aldrich (St. Louis, Mo.).

The kinase reactions are conducted in clear-bottom 96-well plate fromThermo Fisher Scientific in a final volume of 24 μL. Inhibitors arefirst diluted serially in DMSO and added to the plate wells before theaddition of other reaction components. The final concentration of DMSOin the assay is 0.5%. The PI3K assays are carried out at roomtemperature in 20 mM MOPS, pH 6.7, 10 mM MgCl₂, 5 mM DTT and CHAPS0.03%. The reaction mixture is prepared containing 50 μM PIP2, kinaseand varying concentration of inhibitors. Reactions are initiated by theaddition of ATP containing 2.2 μCi [γ-³³P] ATP to a final concentrationof 1000 μM. The final concentration of PI3K isoforms α, β, δ and γ inthe assay were 1.3, 9.4, 2.9 and 10.8 nM, respectively. Reactions areincubated for 180 minutes and terminated by the addition of 100 μL of 1M potassium phosphate pH 8.0, 30 mM EDTA quench buffer. A 100 μL aliquotof the reaction solution are then transferred to 96-well MilliporeMultiScreen IP 0.45 μm PVDF filter plate (The filter plate is prewettedwith 200 μL 100% ethanol, distilled water, and 1 M potassium phosphatepH 8.0, respectively). The filter plate is aspirated on a MilliporeManifold under vacuum and washed with 18×200 μL wash buffer containing 1M potassium phosphate pH 8.0 and 1 mM ATP. After drying by aspirationand blotting, the plate is air dried in an incubator at 37° C.overnight. Packard TopCount adapter (Millipore) is then attached to theplate followed with addition of 120 μL Microscint 20 scintillationcocktail (Perkin Elmer) in each well. After the plate sealing, theradioactivity of the product is determined by scintillation counting onTopcount (Perkin-Elmer). IC₅₀ determination is performed by fitting thecurve of percent control activity versus the log of the inhibitorconcentration using the GraphPad Prism 3.0 software.

Example A3 PI3Kδ Scintillation Proximity Assay Materials

[γ-³³P] ATP (10 mCi/mL) was purchased from Perkin-Elmer (Waltham,Mass.). Lipid kinase substrate, D-myo-Phosphatidylinositol4,5-bisphosphate (PtdIns(4,5)P2)D (+)-sn-1,2-di-O-octanoylglyceryl,3-O-phospho linked (PIP2), CAS 204858-53-7, was purchased from EchelonBiosciences (Salt Lake City, Utah). PI3Kδ (p110δ/p85α) was purchasedfrom Millipore (Bedford, Mass.). ATP, MgCl₂, DTT, EDTA, MOPS and CHAPSwere purchased from Sigma-Aldrich (St. Louis, Mo.). Wheat GermAgglutinin (WGA) YSi SPA Scintillation Beads was purchased from GEhealthcare life sciences (Piscataway, N.J.).

The kinase reaction was conducted in polystyrene 384-well matrix whiteplate from Thermo Fisher Scientific in a final volume of 25 μL.Inhibitors were first diluted serially in DMSO and added to the platewells before the addition of other reaction components. The finalconcentration of DMSO in the assay was 0.5%. The PI3K assays werecarried out at room temperature in 20 mM MOPS, pH 6.7, 10 mM MgCl₂, 5 mMDTT and CHAPS 0.03%. Reactions were initiated by the addition of ATP,the final reaction mixture consisted of 20 μM PIP2, 20 μM ATP, 0.2 μCi[γ-³³P] ATP, 4 nM PI3Kδ. Reactions were incubated for 210 min andterminated by the addition of 40 μL SPA beads suspended in quenchbuffer: 150 mM potassium phosphate pH 8.0, 20% glycerol. 25 mM EDTA, 400μM ATP. The final concentration of SPA beads was 1.0 mg/mL. After theplate sealing, plates were shaken overnight at room temperature andcentrifuged at 1800 rpm for 10 minutes, the radioactivity of the productwas determined by scintillation counting on Topcount (Perkin-Elmer).IC₅₀ determination was performed by fitting the curve of percent controlactivity versus the log of the inhibitor concentration using theGraphPad Prism 3.0 software. IC₅₀ data for the Examples is presented inTable 2 as determined by Assay A3.

TABLE 2 PI3Kδ Example # IC₅₀ (nM)* 1 + 1A + 1B ++ 1C + 1D ++ 2 + 2A + 2B++ 2C + 2D ++ 3 + 4 + *column symbols (for Table 2): + refers to ≤100 nM++ refers to >100 nM to 400 nM

Example B1 B Cell Proliferation Assay

To acquire B cells, human PBMC are isolated from the peripheral blood ofnormal, drug free donors by standard density gradient centrifugation onFicoll-Hypague (GE Healthcare, Piscataway, N.J.) and incubated withanti-CD19 microbeads (Miltenyi Biotech, Auburn, Calif.). The B cells arethen purified by positive immunosorting using an autoMacs (MiltenyiBiotech) according to the manufacture's instruction.

The purified B cells (2×10⁵/well/200 μL) are cultured in 96-wellultra-low binding plates (Corning, Corning, N.Y.) in RPMI1640, 10% FBSand goat F(ab')2 anti-human IgM (10 μg/ml) (Invitrogen, Carlsbad,Calif.) in the presence of different amount of test compounds for threedays. [³H]-thymidine (1 _(μ)tCi/well) (PerkinElmer, Boston, MA) in PBSis then added to the B cell cultures for an additional 12 hours beforethe incorporated radioactivity is separated by filtration with waterthrough GF/B filters (Packard Bioscience, Meriden, Conn.) and measuredby liquid scintillation counting with a TopCount (Packard Bioscience).

Example B2 Pfeiffer Cell Proliferation Assay

Pfeiffer cell line (diffuse large B cell lymphoma) are purchased fromATCC (Manassas, Va.) and maintained in the culture medium recommended(RPMI and 10% FBS). To measure the anti-proliferation activity of thecompounds, the Pfeiffer cells are plated with the culture medium (2×10³cells/well/per 200 μl) into 96-well ultra-low binding plates (Corning,Corning, N.Y.), in the presence or absence of a concentration range oftest compounds. After 3-4 days, [³H]-thymidine (1 μCi/well)(PerkinElmer, Boston, Mass.) in PBS is then added to the cell culturefor an additional 12 hours before the incorporated radioactivity isseparated by filtration with water through GF/B filters (PackardBioscience, Meridenj, Conn.) and measured by liquid scintillationcounting with a TopCount (Packard Bioscience).

Example B3 SUDHL-6 Cell Proliferation Assay

SUDHL-6 cell line (diffuse large B cell lymphoma) was purchased fromATCC (Manassas, Va.) and maintained in the culture medium recommended(RPMI and 10% FBS). To measure the anti-proliferation activity of thecompounds through ATP quantitation, the SUDHL-6 cells was plated withthe culture medium (5000 cells/well/per 200 μl) into 96-well polystyreneclear black tissue culture plate (Greiner-bio-one through VWR, N.J.) inthe presence or absence of a concentration range of test compounds.After 3 days, Cell Titer-GLO Luminescent (Promega, Madison, Wis.) cellculture agent was added to each well for 10 minutes at room temperatureto stabilize the luminescent signal. This determines the number ofviable cells in culture based on quantitation of the ATP present, whichsignals the presence of metabolically active cells. Luminescence wasmeasured with the TopCount 384 (Packard Bioscience through Perkin Elmer,Boston, Mass.). IC₅₀ data for the Examples is presented in Table 3determined by the assay of Example B3.

TABLE 3 SUDHL6 Example # IC₅₀ (nM) 1A + 1C + 2A + 2C + 3 + 4 + * columnsymbols (for Table 3): + refers to ≤500 nM

Example C Akt Phosphorylation Assay

Ramos cells (B lymphocyte from Burkitts lymphoma) are obtained from ATCC(Manassas, Va.) and maintained in RPMI1640 and 10% FBS. The cells (3×10⁷cells/tube/3 mL in RPMI) are incubated with different amounts of testcompounds for 2 hrs at 37° C. and then stimulated with goat F(ab′)2anti-human IgM (5 μg/mL) (Invitrogen) for 17 minutes in a 37° C. waterbath. The stimulated cells are spun down at 4° C. with centrifugationand whole cell extracts are prepared using 300 μL lysis buffer (CellSignaling Technology, Danvers, Mass.). The resulting lysates aresonicated and supernatants are collected. The phosphorylation level ofAkt in the supernatants are analyzed by using PathScan phospho-Akt1(Ser473) sandwich ELISA kits (Cell Signaling Technology) according tothe manufacturer's instruction.

Example J In Vitro JAK Kinase Assay

The compounds in Table 1 were tested for inhibitory activity of JAKtargets according to the following in vitro assay described in Park etal., Analytical Biochemistry 1999, 269, 94-104. The catalytic domains ofhuman JAK1 (a.a. 837-1142), JAK2 (a.a. 828-1132) and JAK3 (a.a.781-1124) were expressed using baculovirus in insect cells and purified.The catalytic activity of JAK1, JAK2 or JAK3 was assayed by measuringthe phosphorylation of a biotinylated peptide. The phosphorylatedpeptide was detected by homogenous time resolved fluorescence (HTRF).IC₅₀s of compounds were measured for each kinase in the 40 μL reactionsthat contain the enzyme, ATP and 500 nM peptide in 50 mM Tris (pH 7.8)buffer with 100 mM NaCl, 5 mM DTT, and 0.1 mg/mL (0.01%) BSA. For the 1mM IC₅₀ measurements, ATP concentration in the reactions was 1 mM.Reactions were carried out at room temperature for 1 hour and thenstopped with 20 μL 45 mM EDTA, 300 nM SA-APC, 6 nM Eu-Py20 in assaybuffer (Perkin Elmer, Boston, Mass.). Binding to the Europium labeledantibody took place for 40 minutes and HTRF signal was measured on aPHERA star plate reader (BMG, Cary, N.C.). The data for the JAK1 and/orJAK2 inhibitors were obtained by testing the compounds in the Example Jassay at 1 mM ATP.

Various modifications of the invention, in addition to those describedherein, will be apparent to those skilled in the art from the foregoingdescription. Such modifications are also intended to fall within thescope of the appended claims. Each reference, including all patent,patent applications, and publications, cited in the present applicationis incorporated herein by reference in its entirety.

1-43. (canceled)
 44. A method of treating a disease selected from Mantlecell lymphoma, follicular lymphoma, extranodal marginal zone lymphoma,and splenic marginal zone lymphoma in a patient in need thereof,comprising administering to said patient a therapeutically effectiveamount of a compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein: Ar is:

R¹ is methyl; R² is C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, or phenyl; whereinsaid phenyl is optionally substituted by 1, 2, or 3 substituentsindependently selected from halo, OH, CN, C₁₋₄ alkyl, C₁₋₄ alkoxy, andC₁₋₄ haloalkoxy; R³ is Cy or C(═O)NR^(c)R^(d); provided that either (i)R² is phenyl, wherein said phenyl is optionally substituted by 1, 2, or3 substituents independently selected from halo, OH, CN, C₁₋₄ alkyl,C₁₋₄ alkoxy, and C₁₋₄ haloalkoxy; or (ii) R³ is Cy; R⁴ is halo, C₁₋₄alkyl, or C₁₋₄ haloalkyl; R⁵ is halo, CN, C₁₋₄ alkyl, or C₁₋₄ haloalkyl;R⁶ is H; each Cy is independently selected from 4-6 memberedheterocycloalkyl or 5-6 membered heteroaryl, each of which is optionallysubstituted with 1 or 2 independently selected R^(3b) groups; each R^(c)and R^(d) is independently selected from H and C₁₋₆ alkyl; and eachR^(3b) is independently selected from halo, CN, C₁₋₆ alkyl, C₁₋₆haloalkyl, hydroxy, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, amino, C₁₋₆alkylamino, di(C₁₋₆alkyl)amino, C₁₋₆ alkylsulfonyl, carbamyl, C₁₋₆alkylcarbamyl, and di(C₁₋₆ alkyl)carbamyl.
 45. The method of claim 44,wherein R² is C₁₋₆ alkoxy or phenyl; wherein said phenyl is optionallysubstituted by 1, 2, or 3 substituents independently selected from halo,OH, CN, C₁₋₄ alkyl, C₁₋₄ alkoxy, and C₁₋₄ haloalkoxy.
 46. The method ofclaim 44, wherein R³ is C(═O)NR^(c)R^(d).
 47. The method of claim 44,wherein R³ is Cy.
 48. The method of claim 44, wherein R⁴ is C₁₋₄ alkylor halo.
 49. The method of claim 44, wherein R⁵ is halo, CN, or methyl.50. The method of claim 44, wherein: Ar is:

R² is C₁₋₆ alkoxy or phenyl, wherein said phenyl is optionallysubstituted by 1, 2, or 3 independently selected halo groups; R³ isC(═O)NR^(c)R^(d), 4-6 membered heterocycloalkyl, or 5-6 memberedheteroaryl; wherein said 4-6 membered heterocycloalkyl and 5-6 memberedheteroaryl is optionally substituted by 1, 2, or 3 independentlyselected R^(3b) groups; each R^(3b) is independently selected from halo,CN, C₁₋₆ alkyl, C₁₋₆ haloalkyl, hydroxy, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,amino, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, C₁₋₆ alkylsulfonyl,carbamyl, C₁₋₆ alkylcarbamyl, and di(C₁₋₆ alkyl)carbamyl; R⁴ is C₁₋₄alkyl or halo; R⁵ is halo; and R⁶ is H.
 51. The method of claim 44,wherein: Ar is:

R² is C₁₋₆ alkoxy; R³ is 4-6 membered heterocycloalkyl; R⁴ and R⁵ areeach independently halo; and R⁶ is H.
 52. The method of claim 44,wherein: Ar is:

R² is C₁₋₆alkoxy or phenyl, wherein said phenyl is optionallysubstituted by 1, 2, or 3 independently selected halo groups; R³ isC(═O)NR^(c)R^(d), 4-6 membered heterocycloalkyl, or 5-6 memberedheteroaryl; wherein said 4-6 membered heterocycloalkyl and 5-6 memberedheteroaryl is optionally substituted by 1, 2, or 3 independentlyselected R^(3b) groups; each R^(3b) is di(C₁₋₆ alkyl)carbamyl; R⁴ ishalo or C₁₋₄ alkyl; R⁵ is halo; and R⁶ is H.
 53. The method of claim 44,wherein the compound is selected from:4-{3-chloro-6-ethoxy-2-fluoro-5-[1-([1,3]thiazolo[5,4-d]pyrimidin-7-ylamino)ethyl]phenyl}pyrrolidin-2-one;4-{3-chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-one;5-{3-chloro-6-methoxy-2-methyl-5-[(1S)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}-N,N-dimethylpyridine-2-carboxamide;4-chloro-N-ethyl-3′,5′-difluoro-3-methyl-6-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]biphenyl-2-carboxamide;or a pharmaceutically acceptable salt thereof.
 54. The method of claim44, wherein the compound is selected from:(S)-4-(3-chloro-6-ethoxy-2-fluoro-5-((S)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl)phenyl)pyrrolidin-2-one;(R)-4-(3-chloro-6-ethoxy-2-fluoro-5-((R)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl)phenyl)pyrrolidin-2-one;(S)-4-(3-chloro-6-ethoxy-2-fluoro-5-((R)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl)phenyl)pyrrolidin-2-one;and (R)-4-(3 -chloro -6-ethoxy-2-fluoro-5-((S)-1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl)phenyl)pyrrolidin-2-one;or a pharmaceutically acceptable salt thereof.
 55. The method of claim44, wherein the disease is Mantle cell lymphoma.
 56. The method of claim44, wherein the disease is follicular lymphoma.
 57. The method of claim44, wherein the disease is extranodal marginal zone lymphoma.
 58. Themethod of claim 44, wherein the disease is splenic marginal zonelymphoma.
 59. The method of claim 44, wherein the compound is4-{3-chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-one,or a pharmaceutically acceptable salt thereof.
 60. The method of claim55, wherein the compound is4-{3-chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-one,or a pharmaceutically acceptable salt thereof.
 61. The method of claim56, wherein the compound is4-{3-chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-one,or a pharmaceutically acceptable salt thereof.
 62. The method of claim57, wherein the compound is4-{3-chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-one,or a pharmaceutically acceptable salt thereof.
 63. The method of claim58, wherein the compound is4-{3-chloro-6-ethoxy-2-fluoro-5-[1-(pyrido[3,2-d]pyrimidin-4-ylamino)ethyl]phenyl}pyrrolidin-2-one,or a pharmaceutically acceptable salt thereof.